褪黑素对结肠黏膜上皮细胞氧化应激的影响

目的 探讨褪黑素对大鼠结肠黏膜上皮细胞氧化应激的影响。方法 在分离培养正常大鼠结肠黏膜上皮细胞基础上，利用FeSO4 H2O2产生*OH攻击制备结肠黏膜细胞氧化损伤模型，同时预先加入褪黑素并观察损伤减轻程度。实验设正常对照组、模型对照组、5-氨基水杨酸给药组(0.1mmol／L)，褪黑素给药组(0．01、0．1、l.0mmol／L)。培养一定时间后，取上清测乳酸脱氢酶(LDH)、丙二醛(MDA)、谷胱甘肽过氧化物酶(GSHPx)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、一氧化氮(NO)和黏液含量。结果 模型组大鼠结肠黏膜上皮细胞：MDA、LDH和NO水平增高，CAT、GSHPX、SOD和黏液含量减少。不同剂量褪黑素呈剂量依赖性的减少LDH释放，抑制MDA和NO的形成，恢复黏液、CAT、GSHPX及SOD水平。结论 褪黑素对大鼠结肠黏膜氧化损伤具有保护作用。     

脂多糖刺激树突状细胞前后TLR2、3、4基因表达的检测

目的 检测脂多糖作用于树突状细胞前后TLR2、3、4基因的表达情况。方法 从外周血分离单核细胞，加入rhGM-CSF及rhIL-4，培养的第6天，实验组加入脂多糖刺激24h，对照组不加。分别提取细胞总RNA，行半定量RT-PCR，产物进行琼脂糖凝胶电泳分析。结果 单核细胞经GM-CSF及IL-4诱导7d后的DC表达较高水平TLR2、3、4。脂多糖刺激24h后，TLR2、3、4表达下降，TLR4几乎测不到。结论 树突状细胞在受脂多糖刺激前后TLRs的表达有显著变化，推测和它的功能成熟有关。     

急性冠脉综合征患者循环内皮祖细胞与炎性相关因子的研究

目的:探讨急性心肌梗死(AMI)、不稳定性心绞痛(UA)患者外周血中循环内皮祖细胞(EPCs)与炎性相关因子C反应蛋白质(CRP)、白细胞介素-6(IL-6)及肿瘤坏死因子-α(TNF-α)的相关性。方法:取AMI(25例)UA患者(28例)及健康体检者(对照组32例)静脉血,采用密度梯度离心法从外周血获得单个核细胞;激光共聚焦显微镜鉴定FITC-UEA-I和Dil-acLDL双染色阳性细胞为正在分化的EPCs,并在倒置荧光显微镜下计数;酶联免疫法检测各种炎性相关因子。结果:AMI组及UA组的EPCs数量和血清CRPI、L-6、TNF-α水平显著高于正常对照组。结论:AMI及UA患者EPCs数量增加与炎性相关因子增加有关。     

ABO血型不合的异基因造血干细胞移植--2例报告附文献复习

探讨ABO血型不合的异基因造血干细胞移植 (Allo -HSCT)的造血问题。方法 :对 2例患者分别进行异基因外周血造血干细胞移植 (Allo -PBSCT)及脐血造血干细胞移植 (CBSCT) ,用血细胞分离机或 6 %羟乙基淀粉去除移植物中的红细胞 ,定期检测受者血型 ,根据情况输注血细胞。结果 :2例患者全部获得造血重建 ,未出现溶血反应。结论 :ABO血型不合可以进行Allo -HSCT ,去除供者移植物中的红细胞能有效地防止急性溶血反应的发生 ,定期检测受者血型 ,及时调整输血方案有重要的临床意义。     

人膀胱癌T24细胞及其SCID小鼠移植瘤组织中KDR的表达

目的 检测KDR在人膀胱癌T24细胞及其SCID小鼠移植瘤组织中的表达。方法 常规培养T24细胞，建立荷人膀胱癌移植瘤SCID小鼠动物模型，使用抗KDR单克隆抗体，通过免疫荧光、免疫组化检测KDR在T24细胞及瘤体组织上的表达；结果 KDR在T24细胞及瘤体组织中均呈阳性表达。结论 本研究结果为进一步研究抗KDR单抗在荷T24细胞移植瘤动物体内分布及对瘤体生长的抑制效应奠定了实验基础。     

Bacterial components regulate the expression of Toll-like receptor 4 on human mast cells

Objectives and design: The aim of this study was to investigate whether the exposure of mast cells (MCs) to bacterial components affects the expression of Toll-like receptor (TLR) 4, and to elucidate the behavior of MCs during the early response to infection. Materials: Two human MC lines, HMC-1 and LAD2, were employed. Messenger RNA expression was observed by RT and real-time PCR. TLR4 expression was determined by Western blotting. TNF-α secretion was analyzed with ELISA. The degranulation ratio was measured with betahexosaminidase assay. Results: Although bacterial components increased TLR4 mRNA, only lipopolysaccharide (LPS) augmented the TLR4 protein expression. LAD2 pre-treated with LPS for 8 h resulted in 2-fold increased TNF-α secretion on LPS restimulation. Conclusion: These results suggest that the exposure of MCs to LPS may reinforce the innate immune system due to up-regulation of MC TLR4, followed by increased TNF-α release. Received 20 April 2006; returned for revision 14 July 2006; accepted by G. Wallace 11 August 2006     

抗肝癌单链双功能抗体在Hut-78细胞系中的表达及体外杀伤活性

目的 用携带分泌型抗肝癌单链双功能抗体基因（sFv-TNF-α）的重组逆转录病毒感染T细胞淋巴瘤Hut-78细胞，使其表达并分泌针对人肝癌细胞的sFv-TNF-α融合蛋白，观察转导的T细胞对体外培养肝癌细胞的杀伤作用。方法 用感染性重组病毒产生细胞C22（PA317/PST）产生的病毒上清转导入T细胞淋巴瘤Hut-78细胞，采用PCR，RT-PCR，细胞原位杂交及免疫组织化学染色等方法，对转导的Hut-78细胞进行DNA，mRNA及蛋白水平的分析。转导的Hut-78细胞分别与SMMC-7721，HHCC共培养，MTT法检测Hut/PST细胞表达产物对两种肝癌细胞的杀伤作用。结果 PCR，RT-PCR结果显示转导Hut细胞中扩增出外源目的基因对应的电泳条带，neo基因探针原位杂交显示转导细胞质内有较强的紫蓝色阳性反应信号，其阳性率约为95%，鼠抗人TNF-α多克隆抗体免疫组织化学染色，转导细胞质内有明确的棕黄色阳性反应信号，MTT法检测结果，分泌型抗肝癌单链双功能抗体对体外培养的两种肝癌细胞的杀伤率分别为15.4%和26.5%。结论 分泌型抗肝癌单链双功能抗体基因可以在T细胞中整合并稳定表达，其分泌的表达产物对两种肝癌细胞均具有一定的亲合活性，并且具有一定的体外杀伤作用。     

Reelin expression is upregulated following ocular tissue injury

Purpose Reelin is important in the guidance of neuronal stem cells in the central nervous system during normal development. We wished to determine whether reelin is expressed in the retina and cornea after injury. Methods Mice underwent laceration of their retina as well as corneal epithelial debridement. The mice were sacrificed at 3 days, and eyes were fixed and stained for reelin expression and reelin messenger ribonucleic acid (mRNA). Results In normal eyes, reelin was expressed only at very low levels in the ganglion cell layer of the retina and the endothelial cell layer of the cornea. In injured eyes, there was marked expression in reelin immunoreactivity in the retina and cornea. Reelin gene expression was seen in the retina and cornea. Conclusions Reelin is expressed during normal retinogenesis. This study shows that reelin is also upregulated following injury to the retina and cornea. The expression of reelin following injury suggests that reelin may play an important role in regulating stem cell trafficking in neuronal and nonneuronal tissues following injury similar to its role in normal organogenesis. For consideration of publication in Graefe’s Archive for Clinical and Experimental Ophthalmology.     

螺旋藻多糖对人宫颈癌Hela细胞体外生长的影响

目的：探讨螺旋藻多糖（PSP）对体外培养的Hela细胞生长的影响。方法：采用MTT法测定 PSP的抗肿瘤活性，并观察其对Hela细胞形态学的影响。结果：随PSP浓度的升高，Hela细胞存活率逐渐降低，抑制率逐渐增加；光镜下观察显示，PSP作用24-48h后，细胞出现明显的形态学改变。结论：PSP能抑制Hela细胞增殖。     

Inhaled allergen-driven CD1c up-regulation and enhanced antigen uptake by activated human respiratory-tract dendritic cells in atopic asthma

Background Dendritic cells (DC) mediate inflammation in rodent models of allergic airway disease, but the role played by human respiratory‐tract DC (hRTDC) in atopic asthma remains poorly defined. Recent data suggest that CD1 antigen presentation by hRTDC may contribute to asthma pathogenesis. Objective To investigate the influence of hRTDC on the balance between atopy and allergic asthma in human subjects and to determine whether CD1 expression by hRTDC is modulated during asthmatic inflammation. Methods Sputum cells were induced from steroid‐naïve, allergen‐challenged and allergen‐naïve subjects (atopic asthmatics, atopic non‐asthmatics and non‐atopic controls). hRTDC were identified using monoclonal antibody labelling and analysis by flow cytometry. Results hRTDC stained HLA‐DR⁺ (negative for markers of other cell lineages) were predominantly myeloid and comprised ∼0.5% of viable sputum cells. Sputum cells were potent stimulators of allogeneic CD4⁺ naïve T cells and enrichment/depletion experiments correlated stimulatory potency with DC numbers. Sputum contained cells that exhibited typical dendritic morphology when analysed by electron microscopy. Myeloid hRTDC were endocytically active, but uptake of FITC‐dextran was enhanced in cells from asthmatics (P<0.001). Despite their increased endocytic capacity, asthmatic myeloid hRTDC appeared mature and expressed increased levels of maturation markers (P<0.05–P<0.001), CD1c, CD1d and langerin (P<0.05). CD1c expression by asthmatic myeloid hRTDC was enhanced upon in vivo allergen challenge (three to ninefold within 24 h; P<0.05). CD11c⁻CD123^high hRTDC were only detected in asthmatic sputum and were increased in number following allergen challenge. Conclusion Despite limited cell numbers, it proved possible to analyse human RTDC in induced sputum, providing evidence that increased antigen uptake and enhanced CD1 presentation by activated hRTDC may contribute to allergic airway disease. CD1 presentation by hRTDC in atopic asthma may therefore constitute a novel target for future intervention strategies.