单用重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白及联合甲氨蝶呤对胶原诱导性关节炎大鼠关节骨破坏影响的实验研究

目的 通过建立胶原诱导性关节炎(CIA)大鼠模型,评价单用重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)及其联合甲氨蝶呤在抑制CIA大鼠关节骨破坏方面的作用及机制.方法 利用皮下注射牛Ⅱ型胶原诱导Wistar大鼠发病,建立CIA大鼠模型.将造模成功,炎症评分≥2分的CIA大鼠随机分为生理盐水组(0.4 ml/周,腹腔注射)、甲氨蝶呤治疗组(1 mg周,腹腔注射)、rhTN FR:Fc治疗组(0.8 mg,每周2次,腹腔注射)、甲氨蝶呤+rhTN FR:Fc治疗组(甲氨蝶呤1 mg/周+rhTNFR:Fc 0.8mg,每周2次,腹腔注射).治疗8周后,处死大鼠,取踝关节拍摄X线片,胫骨上段行微计算机断层扫描技术扫描和制作硬组织切片,观察各组踝关节骨破坏情况,评价胫骨上段骨小梁变化及骨量变化.统计学处理采用SNK-q检验.结果 治疗8周后,rhTN FR:Fc组,甲氨蝶呤+rhTNFR:Fc组骨小梁面积百分数[(29.1±0.3)％,(26.7±0.6)％]及骨小梁数量(4.4±0.5)/mm,( 4.0±0.6 )/mm]明显高于0.9％氯化钠注射液组和甲氨蝶呤组[(12.9±0.5)％,( 13.2±0.4)％与(2.0±0.3 )/mm,(2.2±0.2)/mm](P＜0.01);rhTNFR:Fc组、甲氨蝶呤+rhTNFR:Fc组骨小梁分离度明显小于0.9％氯化钠注射液组和甲氨蝶呤组(P＜0.01).结论 单用rhTNFR:Fc及联合甲氨蝶呤均具有明显抑制关节骨破坏的作用,且其抑制炎症关节周围骨量减少的作用与抑制局部骨小梁数量减少及骨小梁分离度的增大相关.     

重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白对类风湿关节炎患者血清IgM- IgG-和IgA-类风湿因子的影响

目的 观察重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc,商品名益赛普)对类风湿关节炎(RA)患者IgM-类风湿因子(RF),IgG-RF,IgA-RF的影响,探讨rhTNFR:Fc治疗RA的免疫学机制.方法 选择华中科技大学同济医学院附属协和医院及武汉市中心医院2007-2008年110例RA患者,采用随机数字表法随机分为rhTNFR:Fc组和甲氨蝶呤组.rhTNFR:Fc组55例,每周2次皮下注射rhTNFR:Fc(25 mg/次),24周.甲氨蝶呤组55例,每周1次口服甲氨蝶呤片,7.5mg/次起,8周内逐步加到15 mg/次,24周.观察药物对IgM-RF、IgG-RF、IgA-RF的影响,临床疗效评价采用28个关节疾病活动度(DAS28)疗效评定标准.组内治疗前后的差异采用配对t检验分析,组间治疗前后的差异采用两样本t检验分析.结果 ①2组患者病情均明显改善,rhTNFR:Fc的IgM-RF降低时间早于甲氨蝶呤组(P＜0.05).②rhTNFR:Fc组血清IgM-RF (29±16) U/ml明显降低(P＜0.05),IgG-RF (145±20) U/ml和IgA-RF(153±34)U/ml明显升高(P＜0.05).③甲氨蝶呤组IgM-RF (44±14) U/ml,IgG-RF (62±14) U/ml和IgA-RF (66±19) U/ml均明显降低(P＜0.05).④对临床指标的分析表明rhTNFR:Fc治疗RA疗效确切.结论 rhTNFR:Fc与甲氨蝶呤均能有效缓解RA的病情.rhTNFR:Fc能显著降低RA患者血清中IgM-RF的水平,而对IgG-RF,IgA-RF水平有升高作用,可能与其治疗RA的免疫学机制有关.     

弥散加权成像及动态增强扫描对老年早期强直性脊柱炎骶髂关节的评价价值

目的探讨弥散加权成像及动态增强扫描对老年早期强直性脊柱炎骶髂关节的评价价值。方法选择老年早期强直性脊柱炎患者60例作为强直性脊柱炎组,老年健康体检者60例作为对照组。强直性脊柱炎组和对照组进行弥散加权成像及动态增强扫描。结果强直性脊柱炎组骶侧和髂侧表观弥散系数(ADC)值均明显高于对照组(P<0.05)。强直性脊柱炎组和对照组骶侧和髂侧时间-强度曲线(TIC)类型比较差异有统计学意义(P<0.05),强直性脊柱炎组骶侧和髂侧TIC以Ⅱ型为主,对照组骶侧和髂侧TIC以Ⅲ型为主。强直性脊柱炎组骶髂关节达峰时间(TTP)、信号增强比率(SER)、最大上升斜率(MSI)、峰值信号强度(SIpeak)均明显高于对照组(均P<0.05)。ADC、TTP、SER、MSI、SIpeak诊断强直性脊柱炎的最大曲线下面积(AUC)分别为0.82、0.74、0.79、0.71、0.84,灵敏度分别为85.46%、67.83%、74.21%、68.42%、81.26%,特异度分别为72.13%、71.23%、67.58%、73.45%、77.43%。结论弥散加权成像及动态增强扫描对老年早期强直性脊柱炎骶髂关节改变有一定的诊断价值,SIpeak的诊断效能最高。     

重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白对炎性关节炎患者关节置换术后恢复的影响

目的 探讨重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)对炎性关节炎患者关节置换术后恢复的影响.方法 回顾分析67例应用rhTNFR:Fc或传统改变病情抗风湿药(DMARDs)治疗的炎性关节炎患者行关节置换术后伤口感染发生例数、伤口愈合时间、炎症期时间(体温≥37.5 ℃)及抗生素应用时间.根据所应用药物分为rhTNFR:Fc组和传统DMARDs组.其中,rhTNFR:Fc组单用rhTNFR:Fc或rhTNFR:Fc联合传统DMARDs;传统DMARDs组单用或联合应用2种或2种以上传统DMARDs.统计学处理根据数据类型选择t检验或非参数检验.结果 67例患者中,rhTNFR:Fc组18例,传统DMARDs组49例.rhTNFR:Fc组1例出现伤口感染,传统DMARDs组0例,差异无统计学意义(P＞0.05).rhTNFR:Fc组炎症期时间为(4±3) d,传统DMARDs组为(3±3)d,差异无统计学意义(P＞0.05).rhTNFR:Fc组伤口愈合时间为(14.0±3.1)d,传统DMARDs组为(14.7±2.9)d,差异无统计学意义(P＞0.05).rhTNFR:Fc组术后抗生素应用时间为(14.8±9.3)d,传统DMARDs组为(10.3±2.7)d,差异有统计学意义(P＜0.05).结论 炎性关节炎患者围手术期应用rhTNFR:Fc不增加关节置换术后伤口感染发生率,不延长伤口愈合时间及炎症期时间.

Abstract：

Objective To investigate the affect of rhTNFR:Fc on the postoperative recovery of patients with inflammatory arthritis after arthroplasty. Methods Patients with inflammatory arthritis undergoing arthroplasty were included and divided into rhTNFR:Fc group (rhTNFR:Fc only or combined with conven-tional DMARDs) and conventional DMARDs group (monotherapy with or combination of conventional DMARDs). We retrospectively analyzed the incidence of postoperative infection, wound healing time, the febrile period (body temperature ≥37.5 ℃) and the duration of antibiotics treatment after arthroplasty. x2 test and t test were used for statistical analysis. Results Sixty-seven patients were included, 18 in the rhTNFR: Fc group and 49 in the conventional DMARDs group. One postoperative infection occurred in rhTNFR :Fc group but none in the DMARDs group. There was no significant difference by Fisher's exact test (P＞0.05). The febrile duration was (4±3) days in the rhTNFR :Fc group and (3±3) days in the conventional DMARDs group, the difference was not statistically significant (P＞0.05). The wound healing time was (14.0±3.1) days in the rhTNFR :Fc group and (14.7±2.9) days in the conventional DMARDs group, which was not statistically different(P＞0.05). The duration of antibiotics treatment after operation was (14.8±9.3) days in the rhTNFR: Fc group and (10.3±2.7) days in the conventional DMARDs group, the difference was statistically significant (P＜0.05). Conclusion Using rhTNFR:Fc during perioperative period in patients with inflammatory arthritis does not increase the risk of infectious complications or extending wound healing time and the febrile duration.     

对比关节腔内注射重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白对脊柱关节炎和类风湿关节炎膝关节炎的疗效

目的 比较单次膝关节腔内注射重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)对脊柱关节炎(SpA)和类风湿关节炎(RA)膝关节炎的疗效差异.方法 入选确诊SpA或RA并伴有至少一侧膝关节肿胀及积液的受试者,X线显示该膝关节无变形、中重度骨破坏及关节间隙明显狭窄,入组前经过常规剂量改善病情抗风湿药(DMARDs)治疗至少6周,于目标膝关节腔穿刺,吸净滑液后注射1次25 mg rhTNFR:Fc.在注射4周后评价疗效和不良事件,主要疗效指标为改良(纽约)特种外科医院(HSS)膝关节评分.采用配对t检验,两样本t检验和秩和检验进行统计学分析.结果 27例SpA和15例RA受试者入选并完成研究.SpA组改良HSS膝关节评分基线值为(66±14)分,注射4周后为(86±11)分,治疗前后比较差异有统计学意义(P＜0.05)；RA组基线值为(64±13)分,注射4周后为(80±9)分,治疗前后比较差异有统计学意义(P＜0.05).SpA组的改良HSS膝关节评分改善率为24.2％(16.5％～41.9％),RA组为22.2％(15.3％～37.7％),2组比较差异无统计学意义(P＞0.05).SpA组膝关节滑膜厚度改善率为31.8％(9.3％～57.3％),RA组为1.5％(-19.3％～25.5％),2组比较差异有统计学意义(P＜0.05).SpA组6例、RA组2例发生了不良事件,无严重不良事件发生.结论 单次膝关节腔内注射rhTN FR:Fc对SpA和RA膝关节炎安全有效,且SpA膝关节滑膜厚度的减轻程度要大于RA膝关节.     

重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白在胶原诱导性关节炎大鼠中的疗效观察及其对骨桥蛋白的影响

目的 观察重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)在胶原诱导性关节炎(CIA)大鼠中对滑膜、关节软骨等的保护作用,并分析其对骨桥蛋白表达的影响.方法 建立CIA模型,第13天开始对治疗组采用rhTNFR:Fc腹腔注射治疗(10 mg/kg,隔日1次),每周测量体质量及踝关节的前后径,第36天处死大鼠,苏木素-伊红(HE)及甲苯胺蓝染色观察踝关节组织病理变化并进行病理评分;酶联免疫吸附试验(ELISA)检测血浆肿瘤坏死因子(TNF)-α和骨桥蛋白水平,免疫组织化学法检测踝关节骨桥蛋白的组织学表达.多组间均数比较采用单因素方差分析.结果 病理评分定量分析结果 显示造模组与治疗组比较,治疗组关节病理评分显著降低(分别为8.2±1.0与4.8±1.4,P<0.05);ELISA 结果 显示造模组平均血浆TNF-α和骨桥蛋白值分别为(713±146)pg/ml,(4.3±0.6)ng/ml,治疗组分别为(68±20)pg/ml,(4.2±0.6)ng/ml,2组间比较,血浆TNF-α值差异有统计学意义(P<0.05),而骨桥蛋白值差异无统计学意义(P=0.688);免疫组织化学显示骨桥蛋白主要表达在滑膜衬里层、软骨表面,造模组与治疗组间比较差异有统计学意义(P<0.05).结论 rhTNFR:Fc能显著减轻CIA大鼠关节及软骨的破坏,并能显著减少骨桥蛋白在关节滑膜的表达,延缓病情的进展.但不能减少骨桥蛋白在外周血浆中的表达,推测其原因可能为骨桥蛋白不直接参与炎症的发展过程,而主要参与骨质的破坏及吸收过程.

Abstract：

Objective To investigate the protection effects of recombinant human tumor necrosis factor-α receptor Ⅱ :IgG Fc fusion protein for injection (rhTNFR:Fc) on rats with collagen-induced arthritis (CIA) and analyze osteopontin (OPN) changes following therapy in order to understand its primary mechanism of action. Methods CIA was induced by bovine Ⅱ collagen (B Ⅱ C) injection. Rats were treated with rhTNFR:Fc from the 13th day after the first injection of B Ⅱ C till the 36th day. The anterior-posterior diameters of ankle joints and weight were measured weekly. The pathological score was evaluated by HE staining and toluidine blue staining. The blood plasma TNF-α and OPN levels were measured by ELISA and the histology expression was evaluated by immuno-histochemistry. Comparisons between groups were performed with one-way ANOVA. Results Quantitative analysis showed pathological score in the model group and treatment group was significantly reduced in joint pathology (8.2±1.0 vs 4.8±1.4, P<0.05). The mean plasma levels of TNF-α and OPN values were (713±146) pg/ml, (4.3±0.6) ng/ml respectively in the model group,but those of the treatment group were (68±20) pg/ml, (4.2±0.6) ng/ml. Serum TNF-α values were significantly different (P<0.05) between the two groups, while no significant difference was found in the value of plasma OPN (P=0.688) between the two groups. rhTNFR:Fc could reduce the cells OPN expression in the interface layer of the synovium and cartilage (P<0.05). Conclusion Pathology scores and ELISA results haveshown that rhTNFR:Fc has good therapeutic efficacy. It can significantly reduce the bone and cartilage damage of CIA mouse model, and can significantly reduce the expression of OPN in the sliding joints, thereby delay disease progression. However, it can not reduce the expression of OPN in the peripheral blood plasma.OPN may be involved in bone destruction and resorption rather than in inflammatory process.     

耻骨联合磁共振成像急性炎性改变在中轴型脊柱关节炎诊断中的价值及临床因素分析

目的评价耻骨联合MRI急性炎性改变与中轴型脊柱关节炎(ax-SpA)临床特点的相关性。方法回顾性分析我院2014年2月至2020年11月住院的112例ax-SpA患者, 将其分为耻骨联合炎+骶髂关节炎组, 耻骨联合炎+非骶髂关节炎组, 非耻骨联合炎+骶髂关节炎组, 非耻骨联合炎+非骶髂关节炎组共4组, 使用组内相关系数(ICC)分析耻骨联合MRI急性炎症与骶髂关节炎的相关性, 应用单因素方差和χ2检验比较4组之间年龄、性别、BMI、吸烟史、症状持续时间、HLA-B27阳性率、CRP、ESR之间的差异。结果 ax-SpA患者耻骨联合MRI急性炎性改变的发生率为35.7%(40/112)。耻骨联合MRI急性炎症与骶髂关节急性炎症之间无相关性(r=-0.06, P=0.559), 69例骶髂关节急性炎症组有24例同时伴有耻骨联合急性炎症, 43例骶髂关节无急性炎症组有16例有耻骨联合急性炎症。骶髂关节无急性炎症时, 耻骨联合急性炎症组的CRP、ESR分别为(49±60)mg/L、(40±19)mm/1 h, 明显高于非急性炎症组(19±22)mg/L、(22±37)mm/1 h(t=2.36...     

CT引导下骶髂关节注射治疗强直性脊柱炎

目的　研究骶髂关节注射治疗强直性脊柱炎 (AS)的方法和疗效。方法　在CT引导下 ,患者取俯卧位 ,将腰穿针插入骶髂关节注入得宝松 (1ml) /醋酸强的松龙 (5 0mg) +2 %普鲁卡因 2ml。结果　 12例AS ,经骶髂关节注射后 ,10例有效 (显效 7例 ,有效 3例 ) ,2例无效 ,总有效率 83% ,下背痛程度越重、本次发作时间越长及对NSAIDs反应越差 ,疗效越好。结论　骶髂关节注射治疗AS是一种有效的方法 ,近期 (3～ 6个月 )疗效肯定 ,选择合适的患者疗效将更好。     

重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白治疗强直性脊柱炎的多中心双盲随机对照临床研究

目的 评价重组人Ⅱ型肿瘤坏死因子受体-抗体融合蛋白(rhTNFR:Fc)治疗活动性强直性脊柱炎(AS)的临床疗效和安全性.方法 本研究前6周为随机、双盲、安慰剂对照临床试验,后6周为开放研究.143例活动性AS患者随机接受6周的每周2次rhTNFR:Fc(25 mg)或安慰剂皮下注射,主要疗效指标为达到ASAS20的患者比例,次要疗效指标包括达到临床显效的患者比例,与基线值相比Bath AS疾病活动指数、Bath AS功能指数、Bath AS测量指数、脊柱痛、夜间痛、脊柱炎症、患者总体评估指数、肌腱端指数、关节肿胀指数改善的状况.结果 rhTNFR:Fc治疗可使患者获得显著改善,6周时68%患者达到治疗反应,而安慰剂组仅28%(P＜0.001);其他各项疗效指标在治疗组也有明显的改善.rhTNFR:Fc耐受性好,最常见的治疗相关的不良反应为注射部位皮肤反应.结论 rhTNFR:Fc的安全性和耐受性好,能迅速减轻AS的症状和体征,控制AS患者的病情活动.     

结缔组织生长因子及I型和Ⅲ型胶原在脊柱关节病骶髂关节组织中的表达

目的 了解脊柱关节病(SpA)患者骶髂关节中结缔组织生长因子(CTGF)、I型胶原、Ⅲ型胶原的表达情况,探讨CTGF在spA关节软骨纤维化、骨化、关节强直中的作用.方法 30例spA患者(17例双侧影像学骶髂关节炎≥Ⅱ级,13例影像学骶髂关节炎Ⅰ级)均接受CT引导下骶髂关节穿刺活检术,取得骶髂关节组织.组织标本均行苏木素一伊红(HE)染色确认存在骶髂关节炎后,通过免疫组织化学染色方法,标记CTGF、Ⅰ型胶原及Ⅲ型胶原的表达情况.统计学方法采用单因素方差分析和t检验.结果 30例SpA患者骶髂关节组织中CTGF主要在血管翳炎症细胞及骨髓细胞的胞质中高度表达,阳性细胞数明显多于正常组织对照组[(57.9±42.4)腐倍视野和(2.7±2.5),高倍视野);Ⅰ型胶原及Ⅲ型胶原明显沉积于骨、部分软骨及韧带,平均吸光度均明显高于对照组(分别为0.298±0.080和0.044±0.024;28.254±41.165和0.105±0.054).结论 SpA骶髂关节中存在CTGF的高表达,Ⅰ型胶原及Ⅲ型胶原的沉积增多,提示CTGF参与了SpA骶髂关节局部胶原沉积、软骨纤维化变性过程,可能在SpA关节软骨纤维化、关节强直中起重要的作用.

Abstract：

Objective To investigate the expression of connective tissue growth factor(CTGF),coll agen I and collagen Ⅲ in sacroiliac joint(SIJ)of patients with spondyloarthropathy(SpA).Methods Thirty patients with SpA,including 17 patients with grade Ⅱ saeroiliitis and 13 patients with grade Ⅰ sacroiliitis,were performed on CT guided needie biopsy of SIJ.After sacroiliitis were confirmed by staining with hematoxylin and eosin in sacroiliac joint tissue sample,immunohistochemical assay was performed to determine the expression of CTGF,collagen Ⅰ and collagen Ⅲ in sacroiliac ioint tissue.Univariate Chi-square test was used for data comparison between multiple groups and t-test was used for two group data comparison.Results Contrast to healthy controls,CTGF were found upexpressed on the cytoplasm of inflammatory cells in pannus and bone marrow of sacroiliac tissue samples of patients with SpA,while collagen I and collagen Ⅲ were found up-expressed in bone,cartilage and ligament tissue[(57.9±42.4)/HP vs(2.7±2.5)/HP P<0.05,0.298±0.080 vs 0.044±0.024 and 28.254±41.165 vs 0.105±0.054.P<0.05 respectively].Conclusion CTGF,collagen Ⅰ and collagen Ⅲ are up-expressed in SIJ of SpA patients.CTGF may play an important role in articular cartilage fibrosis and ossification of SpA.     

Correlation of serum MMP3 and other biomarkers with clinical outcomes in patients with ankylosing spondylitis: a pilot study

The studies aimed to assess a set of biomarkers for their correlations with disease activity/severity of patients with ankylosing spondylitis (AS). A total of 24 AS patients were treated with etanercept and prospectively followed for 12 weeks. Serum levels of TNF-α, IFN-γ, TGF-β, IL6, IL15, IL17, MMP3, and MICA were measured at baseline and after treatment. The change of these biomarkers was analyzed for correlations with MRI indices for joint inflammation, Bath Ankylosing Spondylitis Disease Activity Index, Bath Ankylosing Spondylitis Functional Index, AS Disease Activity Score, serum CRP, and ESR. The Wilcoxon rank sum test was used to compare the biomarker levels between pre- and post-treatment and between pre-treatment and controls. Both step-wise procedures based on the Akaike information criterion (AIC) and least absolute shrinkage and selection operator with fivefold cross-validation were used to select the best model for pairwise correlations between the above clinical measures and the serum biomarkers. Serum levels of both MMP3 and IL6 were significantly higher in AS patients at baseline. After treatment, the levels of MMP3 decreased, but TGF-β and TNF-α increased significantly?. The changes of serum MMP3 and MICA were significantly associated with MRI sacroiliac joint (SIJ) scores. CRP was positively correlated with serum MMP3 and IL6. The pattern of combined changes of serum MICA, MMP3, TGF-β, IL17, TNF-α, and IFN-γ predicted the MRI score of SIJ by logistic regression analysis. Specific serum biomarkers were significantly associated with clinical measures of AS. Most prominently, serum MMP3 level was found to have a positive correlation with the MRI score of SIJ and CRP. Serum MICA level negatively correlated with disease remission.     

依那西普治疗降低强直性脊柱炎外周血T细胞活性

目的 研究肿瘤坏死因子(TNF)-α抑制剂依那西普(Etanercepl)对强直性脊柱炎(AS)患者外周血T细胞活性的影响.方法 10例健康志愿者,40例活动性AS患者,随机给予依那西普(50 mg,皮下注射,每周1次)或安慰剂治疗,治疗前后分离外周血单个核细胞(PBMC),酶联免疫斑点法(ELISPOT)分别检测分泌TNF-α、白细胞介素(IL)-2、干扰素(IFN)-γ的细胞数量.WST-1法检测T细胞增殖.结果 依那西普治疗后,分泌TNF-α的单核细胞数量减少;抗CD3和抗CD28抗体刺激后,分泌IL-2和IFN-γ的T细胞数量减少.CD4+/CD8+T细胞增殖没有明显变化.结论 抗TNF-α的治疗降低了AS患者外周血T细胞的活性,改善了AS患者病情.     

强直性脊柱炎骶髂关节内皮质激素注射疗效观察

目的了解强直性脊柱炎(AS)骶髂关节(SIJ)内皮质激素注射的疗效.方法患者取俯卧位,在CT导引下穿刺SIJ滑膜部,取组织标本后,在CT导引下原路进入SIJ,每侧注入丙酮缩去炎舒松15mg.记录分析病人术前、术后临床资料.结果28例中22例(78.6%)疼痛明显减轻,晨僵、夜间痛、SIJ压痛、"4”征、ESR升高及需使用NSAIDs的例数均较术前明显减少.所有SIJ标本均有炎症表现.结论SIJ关节内注射长效激素,可作为对NSAIDs不耐受或无效的AS和uSpA病人的一种有效的缓解腰背痛手段,同时uSpA病人SIJ病理标本的获得,有助于早期诊断.     

胰岛素对大鼠心肌梗死后心室重构和心脏功能的影响及其机制

目的:探讨胰岛素对大鼠心肌梗死(MI)后心室重构和心脏功能的影响及其机制。方法: 80只成年雄性Sprague-Dawley大鼠行冠状动脉左前降支(LAD)结扎制备MI模型,随机分为5组:即假手术(Sham)组(n=20)、生理盐水对照(MI+NS)组(n=20)、胰岛素治疗(MI+Ins)组(n=20)、肿瘤坏死因子α(TNF-α)拮抗剂益赛普治疗(MI+En)组(n=10)及Ins+En治疗(MI+Ins+En)组(n=10)。用ELISA法检测各组大鼠在MI后1周和4周时,心肌及血清TNF-α的水平。超声心动图测定各组大鼠左室射血分数(EF)、缩短分数(FS)和左心室舒张末内径(LVEDD)、左心室收缩末内径(LVESD)、经右颈总动脉插管测定血压(BP)、左室舒张压(LVDP)和最大左室舒张压/收缩压变化速率(±LVdp/dtmax）。结果: 大鼠MI后心肌中TNF-α增加,Ins治疗可明显降低大鼠心肌中TNF-α的含量(P<0.05,n=6)。Ins治疗组大鼠EF、FS、LVDP和±LVdp/dtmax均明显高于对照组(P<0.05,n=10),LVESD明显低于对照组(P<0.05,n=10)。与单独En治疗组相比,Ins+En治疗组大鼠EF、FS、LVDP和±LVdp/dtmax明显升高、LVESD明显降低(P<0.05,n=10)。结论: Ins可抑制MI后心室的扩张,改善心脏功能,但其机制不依赖于抑制心肌TNF-α的产生。     

脊柱关节病患者骶髂关节细针活检的病理表现及其临床意义

目的了解骶髂关节(SIJ)炎的病理表现,提高强直性脊柱炎(AS)的早期诊断水平。方法对96例脊柱关节病(SpA)患者的SIJ进行CT导引下的细针活检,3例非SpA死亡病例为对照组。记录临床资料进行分析。结果76例SpA的SIJ存在炎症表现,包括骨髓炎、血管翳形成和炎性细胞浸润,软骨下骨板破坏,软骨变性、破坏,滑膜炎,附着点炎等。其中45例0～Ⅰ级CT下SIJ炎均存在骨髓炎、软骨下骨板炎、软骨变性等改变;0级CT下SIJ炎的滑膜无病理改变,而部分Ⅰ级和所有≥Ⅱ级SIJ炎滑膜可见炎症;软骨破坏和骨化发生率以0～Ⅰ级最低,且仅见于软骨下骨板侧,关节面侧未见破坏;骨质硬化也是0～Ⅰ级最少见;附着点炎仅见于部分≥Ⅱ级SIJ炎;炎性细胞浸润程度Ⅳ级最低。45例0～Ⅰ级CT下SIJ炎的SpA经病理检查诊断为AS;其平均病程显著短于≥Ⅱ级者。结论0级CT下SIJ炎时SIJ已可有炎症存在。SIJ炎可能自骨髓开始,继而血管翳形成,软骨下骨板破坏,软骨变性、破坏,最后纤维化、骨化而导致关节融合。滑膜炎和附着点炎不是SIJ炎的最早改变。病理检查有利于AS的早期诊断和鉴别诊断。     

内皮素在原发性胆汁性肝硬化中作用的初步研究

目的 通过观察原发性胆汁性肝硬化(PBC)患者外周血清内皮素(ET)水平以及应用熊去氧胆酸(UDCA)治疗前后血清中ET的变化,并与肝功能指标进行相关性分析,寻找ET与PBC疾病进展的关系.通过ET作用于外周血单个核细胞(PBMC)引起其分泌的肿瘤坏死因子(TNF)-α及白细胞介素(IL)-6的变化,探讨ET在PBC发病机制中可能的作用.方法 用酶联免疫吸附试验(ELISA)法测量PBC患者外周血清ET的浓度.用ET-1刺激患者PBMC,检测分泌细胞因子的变化.应用反转录-聚合酶链反应(RT-PCR)判断TNF-α基因表达的改变.结果 24例PBC患者血清ET水平为(6.0±2.8)pg/ml,明显高于健康对照组的(2.3±3.4)pg/ml.UDCA治疗前患者ET水平为(7.5±1.6)pg/ml,治疗后为(6.2±2.7)pg/ml,在90%可信区间内具有统计学意义(P=0.084).血清ET水平与患者的肝功能生化指标无明显相关性.ET-1刺激后PBC患者TNF-α的产生有明显增加(P<0.05),IL-6无显著性改变.RT-PCR证实,ET-1刺激PBMC引起TNF-α mRNA的表达增加.结论 PBC患者外周血清中ET浓度明显高于健康志愿者,且可在UDCA治疗后下降.ET-1作用于人PBMC可引起TNF-α mRNA表达上调,TNF-α分泌增加,进而可能激活前炎症因子的级联放大反应,导致PBC疾病的发生发展.     

细辛脑注射液雾化吸入对AECOPD痰中白介素8、肿瘤坏死因子α水平变化的影响

目的 探讨细辛脑注射液雾化吸入配合常规疗法对慢性阻塞性肺疾病急性加重期(acute exacerbation of chronic obstructive pulmonary disease,AECOPD)痰中白介素8(interleukin-8,IL-8)、肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)水平变化的影响.方法 选择住院的AECOPD患者154例,随机分为常规疗法组和联用细辛脑治疗组;联用细辛脑治疗组在常规疗法的基础上联用细辛脑注射液雾化吸入治疗,用ELISA法分别检测其血清及痰液IL-8及TNF-α的含量.结果 常规疗法IL-8含量明显高于联用细辛脑组(3.081 2±0.378 1 vs 1.246 6±0.604 9,P＜0.05);常规疗法TNF-α含量明显高于联用细辛脑组(0.205 5±0.516 6 vs 0.115 2±0.073 4,P＜0.05).结论 在常规疗法的基础上联用细辛脑注射液雾化吸入治疗时可减少AECOPD痰中IL-8、TNF-α的含量.     

类风湿关节炎患者程序化死亡基因5与肿瘤坏死因子-α的表达及其相关性分析

目的 检测类风湿关节炎(RA)及骨关节炎患者血清、滑液中程序化死亡基因(PDCD)5及肿瘤坏死因子(TNF)-α的表达水平,并分析PDCD5与TNF-α表达的相关性,初步探讨PDCD5在RA发病机制中的作用.方法 选取2009年12月至2010年8月50例患者(其中RA 26例,骨关节炎24例),采用酶联免疫吸附试验(ELISA)法检测血清、滑液中PDCD5及TNF-α的含量,统计学分析采用t检验及Pearson直线相关分析.结果 血清中,RA患者的PDCD5含量显著高于骨关节炎患者[(37±33)与(13±14) pg/ml,P＝0.02];滑液中,RA患者的PDCD5含量也显著高于骨关节炎患者[(37±26)与(11±7)pg/ml,P＜0.01].TNF-α在RA患者血清中的含量与骨关节炎患者比较差异无统计学意义(P=0.122),但其在RA患者滑液中的含量显著高于骨关节炎患者(P=0.037).PDCD5与TNF-α在RA及骨关节炎患者的血清中均呈负相关(r=-0.55,P=0.004;r=-0.51,P=0.012),PDCD5与TNF-α在RA患者的滑液中也呈负相关(r=-0.49,P=0.012),但在骨关节炎患者的滑液中无相关(r=-0.353,P=0.09).结论 PDCD5与TNF-α是RA重要的凋亡调控因子,在RA的发生发展过程中发挥重要作用.     

过氧化物酶体增殖物激活受体-γ对干燥综合征非肥胖型糖尿病小鼠的抗炎作用

目的 探讨过氧化物酶体增殖物激活受体(PPAR)-γ对干燥综合征(SS)动物模型非肥胖型糖尿病(NOD)小鼠的抗炎作用.方法 8周龄NOD小鼠20只,随机分为2组,每组10只.实验组自9周龄给予罗格列酮按40 mg/kg剂量隔天灌胃.分别在12周龄和15周龄时随机处死罗格列酮组和对照组各1只小鼠,17周龄时处死全部小鼠,留取外周血和唇腺组织.用苏木素-伊红(HE)染色评价唇腺的病理学改变.酶联免疫吸附试验(ELISA)法测定外周血中白细胞介素(IL)-1β、IL-4、IL-6和肿瘤坏死因子(TNF)-α的含量.实时荧光定量反转录-聚合酶链反应(real-time PCR)检测唇腺组织中IL-1β、IL-4、IL-6和TNF-α mRNA的表达量.计量资料两样本间比较采用t检验.结果 罗格列酮组小鼠唇腺组织中单核细胞浸润较对照组均减轻,腺体破坏少.17周龄时罗格列酮组与对照组比较,外周血IL-6[(26±7)与(37±11),t=-2.298]和TNF-α[(57±22)与(79±21),t=-2.188]的含量明显减少(P<0.05);IL-4[(26±13)与(12±4),t=2.438]的含量明显增加(P<0.05);唇腺组织中TNF-α mRNA的表达水平明显降低;而IL-4 mRNA的表达水平明显升高(P<0.05).结论 PPAR-γ对NOD小鼠干燥综合征有疗效,其作用机制可能与其下调SS体内Th1细胞因子、使Th1/Th2平衡向Th2方向转化相关.

Abstract：

Objective To investigate the anti-inflammatory effect of peroxisome proliferator-activated receptor-γ(PPAR-γ)in non-obese diabetic mice(NOD mice)with Sj(o)gren's syndrome(SS)．Methods Twenty 8-weeks-old female NOD mice were randomly divided into 2 groups．Rosiglitazone and normal saline were administered for the rosiglitazone group and control group respectively．At age of 12 weeks and 15 weeks,one mouse in the rosiglitazone group and the control group were killed respectively,and the others were sacrificed at the age of 17 weeks．Blood were obtained by cardiac puncture,and minor salivary glands (MSG) were resected．The histopathological changes waere examined by H&E staining．The level of IL-1β,IL-4,IL-6 and TNF-α in serum were measured by ELISA．Real-time PCR waft used to evaluate the mRNA expression level of IL-1β,IL-4,IL-6 and TNF-α in MSG．Student's t-test was used to assess the differences．Results Compared with the control group,the mice in the rosiglitazone group showed that：①histopatho-logical change was significantly ameliorated．②At the age of 17 weeks,IL-6[(26±7)vs(37±11),t=-2．298] and TNF-α[(57±22)vs(79±21),t=-2．188] were expressed significantly lower and IL-4[(26±13)vs(12±4),t=2．438 ] was expressed significantly higher in serum(P<0．05)．③The expression of TNF-αwas significantly decreased and the expression of IL-4 was significantly increased in MSG (P<0．05)．Conclusion PPAR-γ can ameliorate SS on NOD mice effectively．The mechanism may be related to reduction of Th1 cytokines,and the Th1/Th2 balance is changed into Th2 predominant．