兔膝关节内注射透明质酸钠后胶原纤维及关节外相关韧带组织的变化

目的:在兔固定的膝关节内定期注入透明质酸钠,观察关节内外组织的变化。方法:实验于2003-10/2004-04在大连医科大学完成。实验分组:新西兰兔24只,随机分为透明质酸钠组、生理盐水组、单纯对照组,每组8只。实验干预:用树脂绷带将兔右膝关节固定于伸直位,膝关节注射部位开窗,左膝自由活动(自身对照侧)。透明质酸钠组膝关节腔内注入透明质酸钠0.1mL,生理盐水组注射同等剂量的生理盐水,单纯对照组不向关节内注射任何药物,每周1次,共5次。实验评估:5周后麻醉下处死动物,去除外固定,采用改良Clarke-Weeknesser关节伸屈范围检查标准测量右膝关节的活动范围;采用改良的Rydell-Balazes肉眼粘连评分标准评估膝关节内纤维粘连情况;光镜下观察股间肌、股直肌、髌内外侧支持带的纤维变性情况。结果:动物饲养过程中死亡4只,进入结果分析透明质酸钠组7只,生理盐水组6只,单纯对照组7只。①右膝关节的活动范围:透明质酸钠组大于生理盐水组和单纯对照组[(37.86±2.94)°,(15.67±2.23)°,(14.29±1.96)°,P<0.01]。②膝关节内纤维粘连评分:透明质酸钠组小于生理盐水组和单纯对照组(1.44±0.49,3.33±0.44,3.44±0.57,P<0.01)。③光镜下透明质酸钠组股间肌、股直肌的纤维变性较生理盐水组和单纯对照组减轻,髌内外侧支持带胶原纤维成分的变化也减轻。结论:在固定的兔膝关节内注射透明质酸钠不但可以明显抑制关节内的纤维性粘连;还可以抑制关节外股间肌、股直肌、髌内外侧支持带的组织变性。     

4例体外受精的胎儿和新生儿发生免疫性血小板减少症的报告

新生儿免疫性血小板减少症(Neonatalalloimmunethrom-bocytopenia,NATP)发病原因是由于母体产生针对胎儿特异性血小板抗原的IgG抗体,并发生抗原抗体反应。胎儿特异性血小板抗原来自父亲。NATP发病率约为0.l%。NATP占新生儿血小板减少症的3%和重度血小板减少(血小板计数<50×109     

Combined plasma exchange and intravenous gammaglobulin in the treatment of patients with refractory immune thrombocytopenic purpura

Plasma exchange (PE) and intravenous gammaglobulin (IVGG) are potentially effective therapies in immune thrombocytopenic purpura (ITP). In this study, eight patients refractory to IVGG and to prednisone were treated with a protocol of combined PE and IVGG therapy using a single PE on each of 3 consecutive days, followed by 2 consecutive days of IVGG at 1 g per kg. Four of the eight patients responded to the combined therapy with a mean peak platelet count of 132,000 per microliter (range, 74,000 to 225,000/microliter). Responses lasted approximately 2 weeks. These four patients were among the five who had initially responded to IVGG before becoming refractory; none of the three patients without an initial response to IVGG responded to the combined therapy. Age, duration of disease, and splenectomy status did not appear to be related to response to the combined therapy. Two patients began maintenance treatment (1 PE followed by 1 g/kg IVGG on the same day), but both became unresponsive after three treatments. PE combined with IVGG may be a useful treatment for some patients with refractory ITP who have uncontrollable bleeding or require major surgery. The development of resistance to IVGG effect may be mediated by an increase in the level of antiplatelet antibodies. PE, by lowering antiplatelet antibody levels, may then allow IVGG infusion to be effective again in elevating the platelet count.     

Autoantibodies against desmoglein 2 are not pathogenic in pemphigus

BackgroundAnti-desmoglein 1 and 3 autoantibodies justify acantholysis in pemphigus; however, the pathogenesis of anti-desmoglein 2 is hypothetical.ObjectiveTo compare the participation of desmogleins 1, 2 and 3 through the production of serum autoantibodies, and protein and gene expression in the skin/mucosa of patients with pemphigus foliaceus and pemphigus vulgaris.MethodsThe autoantibodies were titrated by ELISA in 202 samples of pemphigus foliaceus, 131 pemphigus vulgaris, 50 and 57 relatives of patients with pemphigus foliaceus and pemphigus vulgaris, respectively, and 114 controls. Protein and gene expressions were determined by immunohistochemistry and qPCR in the skin/mucosa of 3 patients with pemphigus foliaceus and 3 patients with pemphigus vulgaris.ResultsHigher titers of anti-desmoglein 2 (optical density) resulted in pemphigus foliaceus and pemphigus vulgaris, when compared to controls (0.166; 0.180; 0.102; respectively; p < 0.0001). There was a correlation between anti-desmoglein 2 and anti-desmoglein 1 titers in pemphigus foliaceus (r = 0.1680; p = 0.0206). There was no cross-reaction of anti-desmoglein 2 with desmoglein 1 and 3. Protein overexpression of desmoglein 2 was observed in intact and lesional skin of patients with pemphigus compared to the skin of controls. Internalization granules of desmoglein 1 and 3, but not of desmoglein 2, were observed in lesions of pemphigus foliaceus and pemphigus vulgaris, respectively. Gene overexpression of desmoglein 2 was observed in the mucosa.Study limitationsSmall sample size for the statistical analysis of protein and gene expression.ConclusionAutoantibodies against desmoglein 2 are not pathogenic in pemphigus; protein and gene overexpression of desmoglein 2 in the skin and mucosa may be involved in acantholysis repair.     

Spinal abnormalities in pediatric patients: MR imaging findings compared with clinical, myelographic, and surgical findings

Eighty-one pediatric patients with a variety of spinal disorders, including suspected dysrhaphism, scoliosis, neoplasia, and neurofibromatosis, underwent magnetic resonance (MR) imaging. The results were retrospectively compared with those of myelography followed by computed tomography (CT) and surgery. In patients with dysrhaphism, most abnormalities, including hydromyelia, inclusion tumors, and sites of cord tether, were demonstrated with MR imaging. Diastematomyelia and small hydromyelic cavities were indistinguishable on routine coronal and sagittal T1-weighted images; axial images with T2 weighting were optimal for this differentiation. MR imaging did not enable direct visualization of a thickened filum or evaluation of tethering with a thin, dorsally positioned neural placode. Congenital or severe scoliosis required lengthy studies with multiple planes of imaging or myelography and CT. Milder curvatures were readily evaluated with MR imaging, and neoplastic lesions, with the exception of intrathecal tumor seeding, were adequately defined.     

应用间断流动式血细胞分离机采集外周血造血干细胞的效果分析

目的:为寻找采集外周血造血干细胞更合适的手段,探讨间断流动式血细胞分离机采集外周血造血干细胞的效果。方法:选择中山大学第五附属医院血液风湿科2004-2006年12例行外周血造血干细胞移植住院患者。①实验对象:供者5例,男1例,女4例,一般状况良好,与患者关系为同胞妹妹3例,同胞弟弟1例,同胞姐姐1例;患者7例,男3例,女4例,23～62岁,7例为自体移植,5例为异基因移植。血液系统疾病患者9例(非霍奇淋巴瘤3例,急性淋巴细胞白血病2例,急性髓细胞白血病2例,慢性粒细胞白血病1例,骨髓增生异常综合征1例);自身免疫性疾病3例(重型系统性红斑狼疮2例,难治复发性类风湿关节炎合并干燥综合征1例)。②实验过程:每位供/患者均知情同意并签署知情同意书。对于自体患者,根据患者的疾病类型采取不同的干细胞动员化疗方案,联合化疗后7～10d,待白细胞下降至最低点,一般为≤1.0×109 L-1时,给予粒细胞集落刺激因子5μg/kg皮下注射,白细胞升至3.0×109 L-1时开始采集;对于allo-PBSCT供者直接给予粒细胞集落刺激因子5μg/kg,皮下注射,1次/d,共5d,同时应用流式细胞仪检测CD34 细胞数,至白细胞升至≥20.0×109 L-1或当CD34 细胞升高>20个/μL时采用增强型多功能血细胞分离机PBSC程序卡采集健康供者和患者外周血造血干细胞。③实验评估:分析采集效率、血液学参数、不良事件发生率、以及供、受者ABO血型不合的患者回输干细胞溶血反应等情况。结果:12例供者、患者均进入结果分析。①共进行了24次采集,其中1次采集1例,2次采集7例,3次采集3例,平均循环次数(25±5)次,采集时间(228±32)min,处理血量(7234±1205)mL,复方枸橼酸钠溶液用量为(623±96)mL,干细胞收集量为(102±21)mL,CD34 细胞采集效率为(54.3±30.7)%,细胞计数示白细胞为(156±34)×109 L-1,单个核细胞为(79.7±13.2)×109 L-1,流式细胞仪检测CD34 细胞为(10.30±4.38)×106/kg受者体质量。②不良反应轻微,24次采集过程中出现不良反应6次均为枸橼酸盐所致低钙血症反应。血红蛋白和血小板与采集前相比分别下降9.36%和11.10%。供、受者ABO血型不合的3例患者在输注造血干细胞悬液后均未出现溶血反应。③12例均获造血功能重建,无移植相关死亡。结论:用间断流动式血细胞分离机采集外周血造血干细胞效果良好,不良反应轻微,能有效的减少被采集者红细胞、血小板的丢失,对供、受者ABO血型不合者不需去除造血干细胞悬液中的红细胞,值得临床应用。     

The changing clinical pattern of Reye's syndrome 1982-1990

OBJECTIVE: To describe trends in the clinical pattern of Reye's syndrome in the British Isles between 1982 and 1990; and to determine the relation between any changes and the June 1986 warnings against the use of aspirin in children. DESIGN: Development, and application to reported cases, of a scoring system designed such that patients showing the typical clinical and pathological features of 'classical' Reye's syndrome scored highly. The relations between 'Reye scores' and a number of explanatory variables were explored using multivariable analysis. SETTING: British Isles. SUBJECTS: 445 cases fulfilling the Reye's syndrome case definition reported to the surveillance scheme between January 1982 and December 1990. MAIN OUTCOME MEASURE: Individual 'Reye score'. RESULTS: Cases with high scores were more likely to have occurred in the 4 1/2 year period before June 1986 compared with the subsequent period (p < 0.006). Numbers of cases in the low and intermediate score categories declined by about 50% after June 1986, whereas those in the high category fell by 79%. High scorers were more likely to have received aspirin (p < 0.0001) and were older than intermediate and low scorers (p < 0.008). No relation was identified between score and season of onset. CONCLUSIONS: The decline in Reye's syndrome after the aspirin warnings cannot be explained entirely, as has been proposed, by improved diagnosis of 'Reye-like' inherited metabolic and other disorders: this would not account for the greater decline of the high scoring subgroup which also contained those cases most likely to resemble 'classical' Reye's syndrome and to have received aspirin. This study provides further evidence for the role of aspirin in a subset of cases meeting the standard diagnostic criteria for Reye's syndrome and supports the need to consider this disorder as a heterogeneous group of conditions including Reye-like inherited metabolic disorders.     

换液频率对骨髓间充质干细胞生物学特性的影响

目的:培养条件是影响骨髓间充质干细胞生物学特性的重要因素。实验考察换液频率对兔骨髓间充质干细胞增殖分化及代谢特性等的影响。方法:实验于2005-03/2005-06在华东理工大学生物反应器工程国家重点实验室完成。①实验材料:1月龄新西兰大白兔购自上海市淞江车墩实验动物良种场。实验过程中对动物处置符合动物伦理学标准。②实验方法:采用密度梯度离心法从兔股骨骨髓中分离得到骨髓间充质干细胞,体外培养扩增后,取生长良好的第3代细胞分别以24,48,72h时间间隔进行换液培养。③实验评估:观察细胞形态学变化,测定细胞生长曲线同时进行乳酸和氨代谢分析,并对几种条件下收获的细胞进行集落形成和成骨分化检测。结果:①每24h换液的细胞最早进入对数生长期,第5天达到增殖顶点,最大细胞数目可达3.44×105,分别是48h和72h换液频率的1.43倍和1.71倍。②每48h换液条件下收获的细胞具有最强的集落形成能力,明显高于每24h和每72h换液条件下收获的细胞。③3种换液频率条件下收获的细胞经成骨诱导后茜素红染色均为阳性,其中每48h换液的细胞胞外钙基质分泌最高。④3种换液频率条件下细胞的代谢曲线无明显差异,乳酸和氨均维持较低浓度,分别在5mmol/L和2mmol/L以下。结论:①换液频率对骨髓间充质干细胞的影响具有双向性。提高换液频率促进骨髓间充质干细胞的增殖,同时也加速了干细胞特性的丢失,导致集落形成能力和成骨分化能力下降。②普遍采用的三四天换液不能提供适合骨髓间充质干细胞生长同时利于干细胞特性维持的营养环境,提示可通过常规培养条件的优化使其有利于骨髓间充质干细胞执行对称的细胞分裂。     

蛋白质组学在白血病中的研究进展

目的:综述蛋白质组学的技术和方法及其在白血病研究中的应用。资料来源:应用计算机检索Medline2001-01/2006-03期间的相关文章,检索词“proteomics、leukemia”,并限定文章语言种类为英文。同时计算机检索维普中文科技期刊数据库2001-01/2006-03期间的相关文章,限定文章语言种类为中文,检索词“蛋白质组、白血病”。并手工查阅相关书籍。资料选择:对资料进行初审,选取近5年的研究进展,筛除重复文献,对剩余的文献开始查找全文。纳入标准:①有关蛋白质组学研究的原著。②有关白血病研究的原著。③有关蛋白质组与白血病关系研究的原著。排除标准:文献中重复研究和Meta分析研究。资料提炼:共收集到107篇有关蛋白质组学与白血病关系研究的文献,排除重复或类似的研究,纳入30篇。其中蛋白质组学方面的有11篇,蛋白质组学与白血病的研究进展的有19篇。资料综合:蛋白质组学是指应用各种科技手段来研究蛋白质组的一门新科学.其目的是从整体的角度分析细胞内动态变的蛋白组成,表达水平与修饰状态。蛋白质组学与基因组学、生物信息学等领域的交叉研究形成了系统的生物学研究模式。白血病是血液系统恶性肿瘤之一,易出现化学治疗后多药耐药,其发生的原因及机制尚不清楚。血液蛋白质组指血液系统中所表达的全部蛋白质,运用蛋白质组分析技术对白血病细胞的增殖、分化与异常转化机制、早期诊断及治疗等方面进行探索,阐明蛋白质表达水平的变化与白血病发生发展的一些相互关系及规律,也为进一步寻找新药和判断预后提供理论依据。即蛋白质组学的发展,对白血病的发病机制、预后判断、化疗药物选择以及个体化治疗带来了重大的意义。结论:将蛋白组学应用于白血病研究,可以对白血病的个体化治疗和预后判断提供理论基础。     

壳聚糖导管复合自体骨髓间充质干细胞修复大鼠13mm坐骨神经缺损

目的:观察壳聚糖神经导管复合自体骨髓间充质干细胞,构建组织工程化人工神经,修复大鼠13mm坐骨神经缺损的效果。方法:实验于2002-10/2004-08在北京军事医学科学院基础医学研究所九室及河北大学附属医院中心实验室完成。实验分组:Wistar大鼠30只按随机数字表法分成3组,实验组、细胞外基质凝胶组和生理盐水对照组,每组10只。壳聚糖神经导管桥接大鼠右侧13mm坐骨神经缺损。实验组无菌条件下抽取股骨髓腔内骨髓组织,贴壁分离法纯化、增殖骨髓间充质干细胞,按1×109L-1细胞浓度与细胞外基质凝胶混合,植入自体神经再生室内;细胞外基质凝胶组神经再生室内植入细胞外基质凝胶,生理盐水对照组神经再生室内植入生理盐水。实验评估:术后12周观察以下指标:①大体观察:观察再生神经。②坐骨神经功能指数测定:选择印迹清晰的足印分别测量正常足(N)和伤侧足(E)的3个指标:足印长度(PL):足尖到足跟的最大距离;足趾宽度(TS):第1～5趾的距离;中间足趾宽度(IT):第2～4趾的距离。结果精确到0.1mm。坐骨神经指数=-38.3[(EPL-NPL)/NPL] 109.5[(ETS-NTS)/NTS]] 13.3[(EIT-NIT)/NIT]-8.8。坐骨神经指数值为0～-11%表示神经功能完全正常,-100%表示神经功能完全丧失,-11%～-100%表示部分神经功能恢复。③电生理检测:检测患侧小腿三头肌肌电图,检测再生神经的运动传导速度和波幅。④腓肠肌湿质量恢复率:腓肠肌湿质量恢复率(%)=手术侧腓肠肌湿质量/对侧正常腓肠肌湿质量×100%。⑤神经组织学检查:苏木精-伊红及Loyez苏木精髓鞘染色,光镜下观察再生神经横断面再生神经髓鞘的形成;Loyez苏木精髓鞘染色及Bielschowsky改良镀银染色,观察纵向切片再生神经神经纤维;神经细丝蛋白免疫组化染色,观察再生的神经轴突染色。随机选取部分正常坐骨神经作为正常对照。⑥透射电镜观察:再生神经的超微结构。⑦再生神经纤维图像分析:观察再生神经有效神经截面积、有髓神经纤维数目、有髓神经纤维密度、有髓神经纤维直径、髓鞘厚度。结果:30只大鼠全部进入结果分析。①大体观察:术后12周,实验组及细胞外基质凝胶组导管内均有再生神经生成,外形似正常神经,直径较正常神经细,生理盐水对照组导管内无再生神经通过间隙。②坐骨神经指数:术后8,12周实验组坐骨神经指数高于细胞外基质凝胶组[分别为(-72.18±3.11)%,(-76.85±2.76)%;(-62.91±2.87)%,(-69.63±2.52)%],差异有显著性意义(F=7.85,P<0.01)。③电生理检查:术后12周实验组运动神经传导速度及波幅明显高于细胞外基质凝胶组[分别为(41.29±3.83),(32.64±3.52)m/s;(3.21±0.34),(2.85±0.22)mV],差异有显著性意义(F=6.39,P<0.01)。实验组、细胞外基质凝胶组腓肠肌肌电图呈部分失神经电位表现,生理盐水对照组则为完全失神经电位。④腓肠肌湿质量恢复率:实验组、细胞外基质凝胶组腓肠肌湿质量恢复率明显高于生理盐水对照组[分别为(69.32±2.65)%,(66.72±1.75)%,(53.41±1.97)%],差异有非常显著性意义(F=9.32,P<0.01),实验组与细胞外基质凝胶组比较差异有显著性意义(F=6.25,P<0.05)。⑤组织学检查:术后12周实验组、细胞外基质凝胶组再生神经纤维排列整齐、密集,神经导管交界处无瘢痕,实验组再生神经纤维多且直径较粗大,排列更为规则。⑥透射电镜观察:实验组和细胞外基质凝胶组均见再生的有髓神经纤维。⑦再生神经纤维图像分析:术后12周实验组有髓神经纤维密度、神经纤维有效面积、有髓神经纤维数量、直径及髓鞘厚度均优于细胞外基质凝胶组。结论:壳聚糖神经导管复合自体骨髓间充质干细胞能够促进周围神经的再生并可以作为种子细胞构建人工神经应用于周围神经缺损的修复。