多发性硬化和格林-巴利综合征患者体液中TNF及IL-1水平与临床关系的研究

测定了多发性硬化(MS)和格林-巴利综合征(GBS)患者的细胞因子，发现活动的MS血清和脑脊液(CSF)TNF水平显著高于稳定的MS和对照组。GBS患者急性期CSF和血清TNF水平显著高于对照组及治疗后水平。另外，MS和GBS患者CSF的TNF水平均高于相应的血清水平。通过研究还发现MS和GBS组CSF的蛋白含量显著增高。此外，MS组CSF白细胞敷与CSF的TNF水平及CSF蛋白含量相关，且CSF蛋白含量与血清、CSF的TNF水平相关。这表明TNF一方面可能来源干鞘内的单个核细胞，另一方面由于血脑屏障受损可能来源于血液，另外还可能来源于神经肢质细胞及血管内皮细胞。TNF可能在炎性脱鞘病发病初期起作用。     

Brain edema and tumor necrosis factor-like weak inducer of apoptosis in rats with cerebral ischemia

BACKGROUND： Recent studies have demonstrated that tumor necrosis factor-like weak inducer of apoptosis （TWEAK） participates in brain edema. However, it is unclear whether blood-brain barrier （BBB） disruption is associated with TWEAK during the process of brain edema OBJECTIVE： To investigate the effects of TWEAK on BBB permeability in brain edema. DESIGN, TIME AND SETTING： An immunohistochemical observation, randomized, controlled animal experiment was performed at the Laboratory of Neurosurgical Anatomy, Xiangya Medical College, Central South University ＆ Central Laboratory, Third Xiangya Hospital, Central South University between January 2006 and December 2007. MATERIALS： A total of 48 adult Wistar rats were randomly divided into three groups： normal control （n = 8）, sham-operated （n = 8）, and ischemia/reperfusion （n = 32）. Rats from the ischemia/reperfusion group were randomly assigned to four subgroups according to different time points, i.e., 2 hours of ischemia followed by 6 hours （n = 8）, 12 hours （n = 8）, 1 day （n = 8）, or 12 days （n = 8） of reperfusion. METHODS： Focal cerebral ischemia/reperfusion injury was induced by middle cerebral artery occlusion （MCAO） using the suture method in rats from the ischemia/reperfusion group. Thread was introduced at a depth of 17-19 mm. Rats in the sham-operated group were subjected to experimental procedures similar to the ischemia/reperfusion group; however, the introducing depth of thread was 10 mm. The normal control group was not given any intervention. MAIN OUTCOME MEASURES： TWEAK expression was examined by immunohistochemistry; brain water content on the ischemic side was calculated as the ratio of dry to wet tissue weight; BBB permeability was measured by Evans blue extravasation. RESULTS： A total of eight rats died prior to and after surgery and an additional eight rats were randomly entered into the study. Thus 48 rats were included in the final analysis. In the ischemia/reperfusion group, TWEAK-positiv     

Cytokine-induced cell death in human oligodendroglial cell lines. II: Alterations in gene expression induced by interferon-gamma and tumor necrosis factor-alpha

Cytokines, such as interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), can initiate dual effects resulting in either cell growth or cell death. In this study, the human oligodendroglial cell lines HOG and MO3.13 were used as a model to study the molecular mechanisms of cytokine-induced cell death in human oligodendrocytes. We have previously shown that TNF-alpha and IFN-gamma induce apoptosis in both oligodendroglial cell lines within 72 hr. In the present study, the cell death pathways operating within these cells were further investigated at the gene expression level. Both cell lines express a broad repertoire of caspases and apoptosis-related genes. Some of these genes are specifically up-regulated by cytokine treatment; e.g., caspase-1 is up-regulated by IFN-gamma. In addition to direct cytotoxic effects, IFN-gamma and TNF-alpha also enhance the expression of Fas, TNFR1, and MHC class I molecules in both cell lines. This suggests that cytokines can make oligodendrocytes more vulnerable to different cell death pathways in an inflammatory environment. cDNA microarray analysis of the HOG cell line revealed that TNF-alpha induces genes that regulate apoptosis, survival, inflammation, cell metabolism, and cell signaling. The data suggest that oligodendroglial cells activate both death and survival pathways upon cytokine challenges. However, the survival pathways seem to be unable to compete with the death signal after more than 24 hr of cytokine treatment. These results may contribute to the development of therapeutic strategies aimed at interfering with cytokine-induced cell death of oligodendrocytes in patients with multiple sclerosis.     

Expression of the costimulatory molecule BB-1 and its receptors in patients with scleroderma-polymyositis overlap syndrome

We investigated expression of costimulatory molecules BB-1, B7-1 (CD80), B7-2 (CD86), and their counter-receptors CD28 and CTLA-4 (CD152) in muscle biopsy specimens of patients with scleroderma-polymyositis overlap syndrome (SSc-PM), primary polymyositis (PM), and other related diseases to examine whether the muscle fibers in patients with SSc-PM behave as antigen-presenting cells (APCs). The major histocompatibility (MHC) class II-positive muscle fibers of SSc-PM patients reacted with monoclonal antibodies (mAb) against BB-1 but not against B7-1 or B7-2. The CD4+ T cells expressed the counter-receptors CD28 and CTLA-4, and bound with the BB-1-positive muscle fibers in cell-to-cell contact. Our findings show that muscle fibers in patients with SSc-PM function as "professional" APCs in a way distinct from muscle fibers in patients with primary PM.     

Increased expression of adhesion molecule CD18 (LFA-1β) on the leukocytes of peripheral blood in patients with acute ischemic stroke

Objectives - The aim of this study was to investigate whether adhesion molecules play a role in acute ischemic stroke. Material and methods -Using immunofluorescence phenotyping and flow cytometry, the expression of leukocyte adhesion molecules CD54, CD11a, CD11b and CD18 in peripheral blood were measured within 12 h after onset of ischemia in 20 patients with stroke. Follow-up measurements were performed at 7 and 30 days after ictus. Results - CD 18 immunofluorescence was significantly increased on the leukocytes within 12 h after onset in patients with stroke compared with the age-matched control group (20 patients with other neurological diseases). Follow-up measurement of CD18 revealed normal results as found in the control group. Conclusion - Our data support the idea that adhesion molecules are involved in tissue injury in ischemic stroke.     

Guillain-Barre综合征患者B淋巴细胞辅助受体CD22、CD72表达的研究

目的探讨Gu illain-Barre综合征(GBS)患者外周血B淋巴细胞辅助受体CD22、CD72表达及其与GBS病程和病情的关系。方法 36例GBS患者(GBS组)按病程及病情分为急性期亚组与恢复期亚组和轻症亚组与重症亚组,应用流式细胞术检测外周血B淋巴细胞CD22及CD72蛋白表达,比较CD22与CD72蛋白在不同亚组间的表达;并与正常对照组(20人)比较。结果 B细胞数量GBS组[(606±118)个]较正常对照组[(248±92)个]明显升高,CD72+CD19+阳性细胞率[(62.11±9.14)%]较正常对照组[(69.72±11.42)%]明显降低(均P<0.01);CD22+CD19+阳性细胞率两组间差异无统计学意义。GBS组中,急性期亚组[(682±91)个]B细胞数量较恢复期亚组[(550±111)个]明显升高,CD72+CD19+阳性细胞率急性期亚组[(57.79±7.69)%]较恢复期亚组[(68.14±7.58)%]明显降低(均P<0.01);CD22+CD19+阳性细胞率两亚组间差异无统计学意义;重症亚组[(685±116)个]较轻症亚组B细胞数量[(561±96)个]明显升高(...     

Circulating tumor necrosis factor (TNF)-α and soluble TNF-α receptors in patients with Guillain-Barré syndrome

Guillain-Barré syndrome (GBS) is an inflammatory disorder that may implicate proinflammatory cytokines such as TNF-α in its pathogenesis. We determined serum levels of TNF-α and the specific antagonists sTNF-Rs p55 and p75 in 24 patients with GBS at days 1, 15 and 30 of hospitalization. Patients were in the progression phase of the disease at day 1, and in the recovery phase at day 30. They were classified as able to walk (stage A), confined to bed (B), or under assisted ventilation (C). All patients underwent plasma exchange within day 1–12. At day 1, TNF-α levels were elevated in

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patients, and sTNF-Rs were elevated in

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. TNF-α levels had not decreased at day 15, and dropped at day 30 (p < 0.04), whereas sTNF-R p55 remained elevated at day 15 and day 30. The TNF-α/sTNF-Rs ratio, estimating active TNF-α unbound to sTNF-Rs, decreased from day 1 to day 30 (p < 0.05). A positive correlation was found between disease severity and sTNF-Rs serum levels (p < 0.01). In conclusion, elevated circulating sTNF-Rs assesses activation of the TNF-α system in almost all patients with GBS and correlates positively with disease severity. Drop of TNF-α contrasting with sustained elevation of sTNF-R p55 during recovery suggests that sTNF-R p55 may be important in the fading of the neural inflammatory effect of TNF-α in GBS.     

肿瘤坏死因子α对大鼠平滑肌细胞增殖及syndecan-4蛋白表达的影响

目的 观察肿瘤坏死因子α（TNF—α）对大鼠胸主动脉血管平滑肌细胞（VSMC）增殖和syndecan-4蛋白表达的影响。方法 体外培养大鼠胸主动脉VSMC，1、10、20、100ng／mL的TNF—α分别作用并设立对照组（不采取干预措施）进行比较，采用MTS／PES法确定VSMC的增殖状态，利用Western blot蛋白免疫印迹法测定syndecan-4蛋白表达。结果 各组细胞增殖率分别为对照组1．001±0．064，TNF—α 1ng／mL组1．082±0．272，10ng／mL组1．125±0．198，20ng／mL组1．566±0．060，100ng／mL组1．215±0．231。统计分析显示低至1ng／mL的TNF-α仍能明显刺激大鼠VSMC的增殖（与对照组比较，P〈0．05），其中以TNF—α 20ng／mL组细胞增殖最显著。TNF-α对VSMC的syndecan-4蛋白的表达有显著的增强作用（与对照组比较，P〈0．05），其中也以20ng／mL组增强作用最显著。结论 TNF-α对体外培养大鼠VSMC增殖和syndecan-4蛋白表达均有显著的促进作用。     

High densities of tumor necrosis factor-α in the cerebral cortex and basal ganglia in human immunodeficiency virus-1-associated cognitive/motor complex: A quantitative regional analysis study

This study determined the regional distribution of tumor necrosis factor-alpha (TNF-α)-positive cells in HIV-1-infected brains and controls, and evaluated its correlation to cognitive impairment in HIV-1-associated cognitive/motor complex (HCMC) using TNF-α and glial fibrillary acidic protein (GFAP) double immunohistochemistry. The TNF-α-positive cell density in the cerebral cortex and basal ganglia was significantly increased in patients with HCMC compared with that in HIV-1-seropositive nondemented patients (non-HCMC), and the basal ganglia TNF-α-positive cell density was significantly correlated with Mini-Mental Status Examination (MMSE) score. Glial fibrillary acidic protein-positive astrocytes were significantly increased in density compared with normal controls, but no significant difference between HCMC and non-HCMC patients was obtained. The density of GFAP-positive astrocytes and TNF-α-positive cells was well correlated other than in the basal ganglia. The basal ganglia TNF-α-positive cell densities showed a significant correlation with MMSE scores. The result suggests that the significantly high content of TNF-α-positive cells in the cortex and basal ganglia is a neurochemical feature characteristic of HCMC. The significant correlation of the basal ganglia TNF-α-positive cell density with degree of cognitive impairment might be associated with subcortical dementia during HIV-1 infection.     

小儿结核性脑膜炎外周血补体因子 H、载脂蛋白A1和淀粉样蛋白A表达水平变化及其临床意义

目的 探讨小儿结核性脑膜炎(Tuberculous meningitis, TBM)外周血补体因子H(Complement factor H,CFH)、载脂蛋白A1(Apolipoprotein AI,ApoAI)和淀粉样蛋白A(Serum amyloid A,SAA)表达水平变化,分析其与TBM病情和预后的关系。方法 选择2017年3月-2021年1月本院收治的112例TBM患儿,根据英国医学研究理事会(Medical research council, MRC)分期标准分为Ⅰ期组(37例)、Ⅱ期组(46例)、Ⅲ期组(29例),根据改良Rankin量表(Modified Rankin Scale, mRS)评分将患儿分为预后良好组(0～2分,62例),预后不良组(≥3分,50例)。另选择71例健康儿童为对照组;检测血清CFH,ApoAI,SAA水平,分析其与TBM病情以及预后的关系。结果 TBM组血清CFH,SAA水平均高于对照组(P<0.05),ApoAI水平低于对照组(P<0.05);重度组血清CFH,SAA水平高于中度组和轻度组(P<0.05),ApoAI水平...     

Increased levels of soluble tumor necrosis factor receptor in patients with multiple sclerosis and HTLV-1-associated myelopathy

Tumor necrosis factor-α (TNF-α) is a potent mediator produced by activated T lymphocytes and macrophages, which may play a role in the pathogenesis and development of multiple sclerosis (MS) and HTLV-1-associated myelopathy (HAM). The first step in the induction of many biological effects elicited by TNF-α is its binding to specific cell surface receptors. A soluble form of TNF receptors (sTNF-R) can be detected in the body fluid. We measured sTNF-R levels in the sera and cerebrospinal fluid (CSF) of patients with either MS or HAM, and evaluated the correlation between this mediator and diseaseaactivity. The levels of sTNF-R in the sera and CSF of patients with MS were significantly increased compared with controls, particularly patients with acute relapsing MS during an exacerbation (P < 0.001). CSF levels of sTNF-R showed a strong correlation with those of TNF (r = 0.716, P < 0.001). Higher levels of sTNF-R in the sera of HAM patients were detected as compared with those of either controls (P < 0.001) or non-HAM carriers (P < 0.001). Patients with HAM exhibited significantly higher CSF levels of sTNF-R than those with other neurological diseases (P < 0.0001). These results suggest that the detection of sTNF-R in the sera and CSF may predict disease progression. Availability of such a marker wou ld be useful in monitoring disease activity.     

干预癫痫大鼠自噬活性对小胶质细胞激活状态及肿瘤坏死因子-α分泌的影响及意义

目的观察干预癫痫大鼠自噬活性对小胶质细胞激活状态及分泌肿瘤坏死因子-α(TNF-α)水平变化的影响,探讨其对神经元以及癫痫状态的影响。方法 Wistar大鼠随机分为正常对照组(6只)与癫痫组(24只);癫痫组大鼠采用戊四氮制作癫痫模型,造模成功后随机分为致痫对照组、3-甲基嘌呤(3-MA)组及雷帕霉素(RAPA)组,每组各6只。观察记录各组大鼠行为学及脑电图变化,采用HE及Nissl染色观察CA1区神经元损伤情况,免疫荧光染色及Western blot检测海马组织LC3、CD68及TNF-α的表达。结果致痫对照组显示癫痫可导致神经元损伤,LC3、CD68、TNF-α的表达较正常对照组显著增加(P0.05)。3-MA组与致痫对照组相比癫痫发作等级降低;神经元损伤数目减少;LC3、CD68、TNF-α的表达显著降低(P0.05)。RAPA组大鼠癫痫发作等级、CD68和TNF-α的表达较致痫对照组无明显变化(P0.05);但神经元损伤数目及LC3的表达进一步增加(P0.05)。结论癫痫过程中存在自噬现象,其可激活小胶质细胞,促进TNF-α分泌,导致神经元损伤;而抑制自噬活性可调控小胶质细胞,减少TNF-α的分泌,保护神经元,从而减轻癫痫发作状态。     

Schwann cell differentiation in Charcot-Marie-Tooth disease type 1A (CMT1A): normal number of myelinating Schwann cells in young CMT1A patients and neural cell adhesion molecule expression in onion bulbs

Charcot-Marie-Tooth disease type 1A (CMT1A) is a common hereditary demyelinating neuropathy caused by a duplication of the gene for the myelin protein PMP22, resulting in overexpression of PMP22 in young patients. Although genetically well defined, the pathogenesis of the hereditary demyelinating neuropathy CMT1A is still unclear. Homology of PMP22 cDNA to the growth arrest-specific gene gas3 and experiments in vitro showing decreased proliferation in PMP22-overexpressing Schwann cells suggest a role of PMP22 in Schwann cell differentiation. Furthermore, overexpression of PMP22 in fibroblasts induces programmed cell death. In this report we applied morphometrical methods using electron micrographs and immunohistochemistry to further characterise Schwann cells in CMT1A nerve biopsy samples from CMT1A patients. We show that the total number of PMP22-expressing Schwann cells, i.e. Schwann cells that are in a 1:1 relationship with axons, was not reduced in sural nerve biopsy samples from six young CMT1A patients. We excluded non-specific secondary Schwann cell proliferation. Thus, in young CMT1A patients with increased PMP22 overexpression there seems to be no evidence for altered initial Schwann cell proliferation in achieving a 1:1 relationship to axons prior to the process of de- and remyelination. Further, using electron microscopy we found no evidence for apoptosis of Schwann cells in CMT1A . However, we provide additional support for an abnormal Schwann cell phenotype in CMT1A by showing the expression of neural cell adhesion molecule immunoreactivity in onion bulbs. Thus, the role of PMP22 in cell growth and differentiation does not lead to an altered number of myelinating Schwann cells but to altered Schwann cell differentiation in CMT1A. Received: 23 September 1996 / Revised: 28 November 1996, 31 January 1997, 2 April 1997 / Accepted: 3 April 1997     

A decrease of human leucocyte antigen-DR expression on monocytes in peripheral blood predicts stroke-associated infection in critically-ill patients with acute stroke

Background and purpose: To investigate changes in human leucocyte antigen (HLA)‐DR expression on peripheral monocytes, determine the value of predicting the development of stroke‐associated infection (SAI), and determine the correlation with other conditions in critically‐ill patients in the neurological intensive care unit (NICU) who suffered an acute stroke. Methods: All patients were enrolled consecutively and admitted to NICU within 24 h after the onset of symptoms. Patients were followed in order to identify whether infection developed and determine survival status within 2 weeks after the stroke. Patients were divided into stroke or control group by study design, infection or non‐infection group by whether or not they had an infection, survival or death group by prognosis and cerebral infarction or cerebral haemorrhage group by stroke type. Patients in which acute stroke was excluded by head CT or MRI were admitted to general ward and were used as a control group. Blood samples were collected serially on days 1, 2, 4, 6 and 14 after stroke, then monocyte human leucocyte antigen‐DR (HLA‐DR) expression was determined by flow cytometry. The National Institute of Health Stroke Scale (NIHSS), Acute Physiology and Chronic Health Evaluation II (APACHEII) and Glasgow Coma Scale (GCS) scores were recorded over the course of observation. Results: Fifty‐three subjects and 39 controls were enrolled in the study. On days 1, 2, 4, 6 and 14, there was a significant difference in monocyte HLA‐DR expression between stroke group and control group (all P < 0.001), but no difference was found between ischaemic stroke group and haemorrhagic stroke group (all P > 0.05). The infection group compared with non‐infection group did not exhibit a significant difference in HLA‐DR expression on days 1 and 2 (all P > 0.05), but significant differences emerged on days 4, 6 and 14 (all P < 0.01). On days 1 and 2 the HLA‐DR expression in the survival group compared with death group, was not significantly different (all P > 0.05), but differences became significant on days 4 and 6 (P < 0.01). On day 1, HLA‐DR expression <62.80% had the predictive value to SAI (sensitivity 83.3%, specificity 55.2%, AUC = 0.661, P = 0.031) and on day 2, HLA‐DR expression <57.83% had the predictive value to SAI (sensitivity 95.8%, specificity 79.3%, AUC = 0.907, P = 0.000) in acute stroke patients. A statistically significant inverse correlation was found between NIHSS and HLA‐DR on days 2 and 4 during the observation period (all P < 0.01), but there was no statistically significant negative correlation on days 1, 6 or 14 (all P > 0.05). HLA‐DR expression did not correlate with APACHEII (all P > 0.05) or GCS (all P > 0.05) during the measurement period. Conclusions: Human leucocyte antigen‐DR (HLA‐DR) expression decreases and sustains a dynamic change and it also relates to the severity of patient’s condition in the critically‐ill patients with stroke. Progressively persistent low monocyte HLA‐DR expression is associated with a poor prognosis. The decline in HLA‐DR expression contributes to infection in critically‐acute stroke patients. Monitoring of monocyte HLA‐DR expression may be useful for identifying patients suffering from acute stroke who are at high risk for infection.     

A case of SUDEP in a patient with Dravet syndrome with SCN1A mutation

A boy with a clinical history of pharmacologically resistant Dravet syndrome died suddenly after falling asleep. The autopsy concluded that the cause of death was sudden unexpected death in epilepsy (SUDEP). Postmortem molecular analysis of the SCN1A gene by multiplex ligation‐dependent probe amplification (MLPA), high‐resolution melting curve analysis (HRMCA), and sequencing revealed a frameshift duplication of adenosine at position 504. The incidence of this mutation is discussed as a potential cause of SUDEP.     

Effect of tumor necrosis factor α and β on human oligodendrocytes and neurons in culture

Cytokines produced by infiltrating hematogenous cells or by glial cells activated during the course of central nervous system disease or trauma are implicated as mediators of tissue injury. In this study, we have assessed the extent and mechanism of injury of human-derived CNS oligodendrocytes and neurons in vitro mediated by the cytokines tumor necrosis factor α and β and compared these with the tumor necrosis factor independent effects mediated by activated CD4⁺ T-cells. We found that activated CD4⁺ T-cells, but not tumor necrosis factor α or β, could induce significant release of lactate dehydrogenase, a measure of cell membrane lysis, from oligodendrocytes within 24 hr. Neither induced DNA fragmentation as measured using a fluorescence nick-end labelling technique. After a more prolonged time period (96 hr), tumor necrosis factor α did induce nuclear fragmentation changes in a significant proportion of oligodendrocytes without increased lactate dehydrogenase release. The extent of DNA fragmentation was comparable to that induced by serum deprivation. Tumor necrosis factor β effects were even more pronounced. In contrast to oligodendrocytes, the extent of DNA fragmentation, assessed by propidium iodide staining, induced in neurons by tumor necrosis factor α was less than that induced by serum deprivation. In-situ hybridization studies of human adult glial cells in culture indicated that astrocytes, as well as microglia, can express tumor necrosis factor α mRNA.     

Detection of tumor necrosis factor-α-positive cells in cerebrospinal fluid of patients with HTLV-I-associated myelopathy

Tumor necrosis factor (TNF)-α-positive cells constituted 1.6–18% and 8.2–23.5% of the total number of cerebrospinal fluid cells from six of 12 patients with HTLV-I-associated myelopathy and in all samples obtained from inflammatory cases, respectively. However, in non-inflammatory cases no TNF-α-positive cells were detected. These results suggest that some of the infiltrating CSF cells produce TNF-α, which plays a role in host immune defenses against causative agents including HTLV-I and in lesion formation within the central nervous system in inflammatory diseases.     

Regulation of monocyte chemoattractant protein-1 expression in adult human non-neoplastic astrocytes is sensitive to tumor necrosis factor (TNF) or antibody to the 55-kDa TNF receptor

Infiltration of the central nervous system (CNS) by monocytes is a characteristics of many non-malignant disease processes, although the signals regulating such traffic are unclear. Tumor necrosis factor (TNF) and other inflammatory cytokines have been shown to elicit production of monocyte chemoattractant activity in glioma cells, but the regulation of such activity in non-neoplastic adult astrocytes has not been examined. We previously observed that TNF constituted a proliferative signal for non-neoplastic adult human astrocytes in vitro involving the 55-kDa TNF receptor. In the present study, we demonstrate that TNF exposure enhances the expression of monocyte chemoattractant protein-1 (MCP-1) mRNA and functional monocyte chemoattractant activity in non-neoplastic astrocytes. Results indicated that MCP-1 mRNA expression was maximal within 3 h, and was further augmented by the protein synthesis inhibitor cycloheximide (CY). Antibody (htr-9) directed against the 55-kDa TNF receptor also elicited MCP-1 mRNA expression while antibody to the 75-kDa TNF receptor (utr-1) was ineffective. Secretion of monocyte chemoattractant activity was significantly greater in TNF- or htr-9-treated astrocytes than in utr-1-treated or untreated controls; activity was abolished by treatment with antibody to MCP-1. These findings suggest that non-neoplastic adult human astrocytes may contribute to CNS inflammatory responses by mediating recruitment of peripheral blood monocytes.