Immunoblotting characterization of neutrophil antigenic targets in autoimmune neutropenia

We characterized neutrophil autoantigens using an immunoblotting technique with antibodies obtained from patients with autoimmune neutropenia. These results were correlated with serologic characterization of the antibodies, using indirect immunofluorescence and leukoagglutination. Of the 17 sera immunoblotted, 16 showed discrete bands in the molecular weight range of 30 to 112. Three patients with Felty's syndrome reacted with an antigenic target of 80 to 84 Kd molecular mass, a finding not seen in any of the other patients studied. By serologic testing, none of the autoimmune sera showed serologic specificity for any known neutrophil-specific alloantigen. Using an anti-NA-1 serum, we identified antigenic targets at 40, 50, and 101 Kd in both NA-1-positive and NA-1-negative neutrophils. Ten of 17 autoimmune sera showed reactivity in this corresponding range. These studies demonstrate that immunoblotting may be used to identify antigenic targets in autoimmune neutropenia and may suggest a specificity of these antibodies not definable by serologic techniques. Correlation of immunoblot reactivity with disease states associated with immune neutropenia may be useful in the study of the pathogenesis of the different forms of autoimmune neutropenia.     

Effect of protein dose on serum glucose and insulin response to sugars

To clarify the effects of protein on insulin and glucose response to sugars, 14 healthy normal-weight males and females were fed test meals containing 0, 15.8, 25.1, 33.6, and 49.9 g protein along with approximately 58 g carbohydrate. Serum samples were obtained at fasting time zero and 15, 30, 60, and 120 min postprandial. Mean areas of the glucose curves above fasting decreased with increasing protein dose. Protein-containing meals produced significantly lower (p less than 0.01) areas than the protein-free meal and the relationship between blood glucose area and protein dose was significant (p less than 0.001). Protein-containing meals produced significantly greater (p less than 0.01) insulin areas compared with the protein-free meal. However, no differences in insulin areas among the protein-containing meals were observed. These data support previous studies showing a blood glucose moderating and insulin-enhancing effect of protein ingestion.     

心肌坏死标记物在检测心肌坏死中的应用与地位

目的:评估各种心肌坏死标记物对判断心肌坏死的价值。资料来源:应用计算机检索LWW全文英文数据库1994-01/2006-08与心肌坏死标记物相关的文献,检索词“Myocardial damage,myocardiolysis,marker”,并限定文章语言种类为“English”。资料选择:对资料进行初审,选对于临床有意义的心肌坏死物进行较详细的总结。选择临床研究的文章,无论有无对照组、是否为随机对照文献均纳入;排除观察对象为动物的基础研究和综述类文献。资料提炼:对收集到的文献进一步查找全文,凡是对于心肌坏死检测快速且可靠的心肌坏死物优先选择,研究内容相似的,以近3年且发表在较权威杂志者优先,最后纳入30篇关于心肌坏死标记物的文章进行综述。资料综合:急性血栓形成及其随后的心肌损伤、坏死是急性心肌梗死的最主要特征,准确、快速、可靠地检测血栓形成和心肌坏死标志物对内科医生诊断急性心肌梗死极具价值。研究表明缺血修饰白蛋白、心肌型脂肪酸结合蛋白、髓过氧化物酶、CK-MB和心肌肌钙蛋白对急性心肌梗死的诊断各具特点,而联合检测更有助于早期、准确诊断急性心肌梗死,为尽早抢救急性心肌梗死创造时机。结论:缺血修饰白蛋白、心肌型脂肪酸结合蛋白、髓过氧化物酶、CK-MB和心肌肌钙蛋白是较敏感的心肌坏死标记物,联合应用诊断价值更大。     

针刺抽吸法诱导建立椎间盘退行性变的动物模型

目的:用针刺抽吸法建立椎间盘退行性变动物模型,分析其可行性和优点。方法:实验于2006-10/2007-03在苏州大学附属第一医院骨科实验室完成。新西兰大白兔18只,用持针器夹持21G皮肤穿刺针从椎间隙正前方刺入L3～4、L4～5、L5～6椎间盘的纤维环,深度控制在5mm,拔出针芯,抽吸出部分髓核组织。术前及术后4,8,12周分别对造模后椎间盘及对照椎间盘(L2～3)行免疫组织化学及组织学观察。结果:新西兰大白兔18只均进入结果分析。①组织学观察:苏木精-伊红染色可见对照组髓核组织中有着大量髓核细胞,造模组第4周髓核细胞数量不断减少,第12周时髓核中主要为纤维软骨组织,伴有极少量髓核细胞。Ⅱ型胶原免疫组织化学检测可见:对照组髓核组织间质呈强阳性染色,而各模型组呈弱阳性染色。随着造模时间的延长,染色逐渐减弱,说明髓核组织中Ⅱ型胶原的含量逐渐减少。②髓核细胞凋亡率:对照组、实验组4,8,12周的髓核细胞凋亡率为(3±1)%,(17±4)%,(36±2)%,(23±5)%,差异有显著性意义(F=113.86,P=0.0001<0.01)。结论:纤维环穿刺吸出部分髓核的方法在不破坏自身免疫屏障的前提下可诱导兔椎间盘的缓慢退行性变,为干细胞、髓核细胞、基因治疗等提供有效的动物模型。     

富碘中药海藻对甲状腺细胞凋亡及凋亡调控基因的影响

目的:通过观察富碘中药海藻对碘缺乏机体甲状腺滤泡上皮细胞凋亡,Fas,FasL,Bcl-2蛋白表达的影响,分析富碘中药过量对甲状腺损伤的机制。方法:实验于2006-03/09在辽宁中医药大学实验中心完成。①实验干预:选用健康鼠龄4周的Wistar大鼠150只。取120只喂低碘饲料建立缺碘大鼠模型,随机分为4组,每组30只:单纯高碘组喂含碘2000μg/L的双蒸水;模型组喂等体积双蒸水;常规剂量海藻组和3倍剂量海藻组分别灌胃海藻生药量13.5g/(kg·d)和40.5g/(kg·d)。以其余30只大鼠为正常对照组:正常饮食,每日灌服等体积双蒸水。②实验评估:分别在给药0,7,28d后取材。采用脱氧核糖核苷酸末端转移酶介导原位缺口末端标记确定甲状腺滤泡上皮细胞凋亡细胞数。采用免疫组化方法观察甲状腺滤泡上皮细胞Fas,FasL,bcl-2表达。结果:大鼠150只均进入结果分析。①凋亡细胞数:给药后7d,常规剂量海藻组和3倍剂量海藻组低于模型组,差异有非常显著性意义(P<0.01)。常规剂量海藻组和3倍剂量海藻组低于单纯高碘组,差异有显著性意义(P<0.05)。给药后28d,正常对照组甲状腺滤泡上皮细胞凋亡细胞数低于其他4组,差异有显著性或非常显著性意义(P<0.05～0.01)。常规剂量海藻组高于单纯高碘组,差异有显著性意义(P<0.05)。②Fas,FasL,Bcl-2蛋白表达:给药7d后,常规剂量海藻组和3倍剂量海藻组Fas蛋白的表达低于模型组,差异有显著性意义(P<0.05)。单纯高碘组和3倍剂量海藻组FasL表达低于模型组,差异有显著性意义(P<0.05)。常规剂量海藻组Bcl-2蛋白表达高于模型组,差异有显著性意义(P<0.05)。给药28d后,单纯高碘组和3倍剂量海藻组Fas蛋白的表达低于模型组,差异有显著性意义(P<0.05)。单纯高碘组、常规剂量海藻组和3倍剂量海藻组Bcl-2蛋白表达高于模型组,差异有显著性意义(P<0.05)。结论:富碘中药海藻给药28d可造成碘缺乏大鼠甲状腺细胞损伤,Fas,FasL,bcl-2可能参与诱导细胞凋亡。     

Complete genomic sequence of Pasteurella multocida, Pm70

We present here the complete genome sequence of a common avian clone of Pasteurella multocida, Pm70. The genome of Pm70 is a single circular chromosome 2,257,487 base pairs in length and contains 2,014 predicted coding regions, 6 ribosomal RNA operons, and 57 tRNAs. Genome-scale evolutionary analyses based on pairwise comparisons of 1,197 orthologous sequences between P. multocida, Haemophilus influenzae, and Escherichia coli suggest that P. multocida and H. influenzae diverged approximately 270 million years ago and the gamma subdivision of the proteobacteria radiated about 680 million years ago. Two previously undescribed open reading frames, accounting for approximately 1% of the genome, encode large proteins with homology to the virulence-associated filamentous hemagglutinin of Bordetella pertussis. Consistent with the critical role of iron in the survival of many microbial pathogens, in silico and whole-genome microarray analyses identified more than 50 Pm70 genes with a potential role in iron acquisition and metabolism. Overall, the complete genomic sequence and preliminary functional analyses provide a foundation for future research into the mechanisms of pathogenesis and host specificity of this important multispecies pathogen.     

Unknown poisoning amongst serving personnel

Background: Poisoning is recognized as an important health problem in many countries of the world. There is incomplete information on poisoning accidents, which is a major problem in developing countries.