环磷酰胺对家兔血液流变性的影响

观察抗肿瘤药物环磷酰胺（ＣＴＸ）对家兔血液流变性的影响。结果显示：ＣＴＸ给药组（１０只）多项血液流变学指标明显异常，与正常对照组（８只）比较具有显著性差异（Ｐ＜０．０５～０．００１），标志着ＣＴＸ有引起高粘滞血症的作用。提示：恶性肿瘤病人应用ＣＴＸ后，可加重原有的高粘滞血症。这一结果可作为肿瘤化疗并用活血化瘀及抗凝治疗的依据     

尿激酶型纤溶酶原激活物系统对骨巨细胞瘤细胞基质金属蛋白酶-2和组织基质金属蛋白酶抑制物-3表达的影响

目的:研究尿激酶型纤溶酶原激活物(uPA)及其受体(uPAR)信号转导对骨巨细胞瘤基质金属蛋白酶-2(MMP-2)和金属蛋白酶组织抑制物-3(TIMP-3)的调节。方法:用免疫组化检测骨巨细胞瘤组织中uPAR、MMP-2和TIMP-3的表达。用免疫共沉淀法检测uPA对瘤细胞信号转导通路的p44蛋白磷酸化水平。用蛋白印迹法检测用uPA和uPAR抗体处理后瘤细胞MMP-2和TIMP-3蛋白表达。结果:(1)uPAR主要表达在部分单核基质细胞和一些多核巨细胞的胞膜上;(2)MMP-2主要表达在瘤细胞的胞浆,在多核巨细胞,其表达有明显的极向性;(3)在骨巨细胞瘤组织TIMP-3表达量低于MMP-2,在多核巨细胞也显示极向性表达;(4)将uPA-ATF加入培养的骨巨细胞瘤细胞后,细胞信号通路上的p44蛋白磷酸化水平明显增高。用uPAR抗体处理后,细胞p44蛋白磷酸化水平明显降低。说明uPA-ATF参与细胞信号转导,而且受uPAR拮抗剂的影响;(5)uPA-ATF信号通路上调MMP-2和TIMP-3的表达,而uPAR抗体则下调MMP-2和TIMP-3的表达。结论:本实验首次直接证明uPA-ATF通过信号转导能调节MMP-2和TIMP-3的表达,而后者则在骨巨细胞瘤局部骨质吸收中起重要作用。     

右旋柠烯抑制小鼠S-180移植瘤生长的实验观察

目的观察右旋柠烯乳剂(D-limonene)对小鼠移植瘤生长及转移的抑制作用。方法用小鼠S180癌性腹水建立皮下移植瘤模型,随机分为3组,预防组建模当天开始给药;治疗组移植后第5天开始给药;对照组不给药。每3d测量肿瘤大小,20d后取材,观察肿瘤细胞的浸润程度及淋巴结转移。结果对照组肿瘤体积较大,与皮肤及深部组织发生粘连,7只发现颈部或腋下淋巴结肿大。治疗组和预防组移植瘤生长速度明显缓慢,体积小,且瘤体较规则,未发现有淋巴结转移者。结论右旋柠烯有抑制肿瘤生长,减少淋巴结转移的作用,其机制有待进一步探讨。     

Sequence analysis of cDNAs for the human and bovine ATP synthase β subunit: mitochondrial DNA genes sustain seventeen times more mutations

We have cloned and sequenced human and bovine cDNAs for the subunit of the ATP synthase (ATP-synß), a nuclear DNA (nDNA) encoded oxidative phosphorylation (OXPHOS) gene. The two cDNAs were found to share 99% amino acid homology and 94% nucleotide homology. The evolutionary rate of ATPsynß was then compared with that of two mitochondrial DNA (mtDNA) ATP synthase genes (ATPase 6 and 8), seven other mtDNA OXPHOS genes, and a number of nuclear genes. The synonymous substitution rate for ATPsynß proved to be 1.9 × 10^–9 substitutions per site per year (substitutions × site^–1 × year^–1) (SSY). This is less than 1/2 that of the average nDNA gene, 1/12 the rate of ATPase 6 and 8, and 1/17 the rate of the average mtDNA gene. The synonymous and replacement substitution rates were used to calculate a new parameter, the selective constraint ratio. This revealed that even the most variable mtDNA protein was more constrained than the average nDNA protein. Thus, the high substitution mutation rate and strong selective constraints of mammalian mtDNA proteins suggest that mtDNA mutations may result in a disproportionately large number of human hereditary diseases of OXPHOS.     

TGFβ1基因对血管内皮细胞细胞外基质表达及与基质黏附的影响

了解TGFβ1基因对血管内皮细胞表达细胞外基质蛋白及与基质黏附力的影响。用DOTAP脂质体转染p MAMneo TGFβ1于原代培养的脐静脉内皮细胞,经G4 18筛选,TGFβ1表达经免疫荧光鉴定。Western blot确定 型胶原、纤黏连蛋白的表达,微管吸吮系统确定内皮细胞与基质的黏附力。结果表明生理情况下的内皮细胞能表达少量的TGFβ1及胶原、纤黏连蛋白。经G4 18筛选,外源性TGFβ1在血管内皮细胞中稳定表达,能显著提高胶原、纤黏连蛋白纤维的表达及细胞与基质的黏附。说明TGFβ1在血管组织工程中促进内皮细胞的黏附具有一定的应用价值。     

IL-10 has a protective role in experimental autoimmune uveoretinitis

The role of IL-10 in the regulation of ocular autoimmune disease was studied in experimental autoimmune uveoretinitis (EAU) elicited in mice by immunization with the retinal antigen interphotoreceptor retinoid binding protein. IL-10-deficient mice were susceptible to EAU, indicating that pathogenesis can occur without presence of IL-10. Treatment of normal mice with IL-10 for 5 days after uveitogenic immunization ameliorated subsequent EAU scores, and down-regulated antigen-specific production of tumor necrosis factor-alpha and IFN- gamma. A concomitant treatment with IL-4 further reduced disease, and resulted in emergence of antigen-specific IL-4 and IL-10 production, as well as in enhancement of the IgG1 antibody isotype. IL-4 by itself was not protective. Only IL-10, but not IL-4, was able to inhibit the function of differentiated uveitogenic T cells in culture. Expression of mRNA for Th1 and Th2 cytokines in the eye during the course of EAU showed that while a Th1 pattern predominated early, IL-10 mRNA expression coincided with down-regulation of the Th1 response and resolution of EAU. Systemic neutralization of IL-10 during the expression phase of EAU resulted in elevated disease scores. Our results suggest that endogenous IL-10 limits expression of EAU and may play a role in the natural resolution of disease. The data further suggest that exogenous IL-10 may be useful in therapeutic control of autoimmune uveitis. While IL-10 by itself is sufficient to suppress Th1 effector development and function, a concomitant administration of IL-4 is required to shift the autoimmune response towards a non-pathogenic Th2 pathway.      

小鼠胃癌MAGE3抗原H-2K~k限制性抗原肽的筛选

本研究对小鼠胃癌MAGE3抗原H 2Kk 限制性的抗原肽进行了筛选。用CTL表位预测的基序法对易与H 2Kk 结合的多肽序列进行了预测并人工合成了 4个多肽。在体外检测了各多肽刺激H 2Kk 型小鼠T细胞增殖和分泌IFN γ的能力 ,同时测定了各多肽诱导出的CTL对小鼠前胃癌细胞株MFC细胞的杀伤活性。结果显示 ,抗原肽MAGE31 3 0 1 3 7可有效地刺激特异性T细胞的增殖和分泌细胞因子IFN γ ,其诱导出的CTL细胞对MFC细胞也有较高的杀伤活性。这些实验结果证实抗原肽MAGE31 3 0 1 3 7可以作为多肽疫苗来对表达MAGE3抗原的H 2Kk 型小鼠胃癌进行免疫治疗。     

糖皮质激素对膝关节周围成骨细胞一氧化氮合酶和骨重建的影响

目的研究糖皮质激素对膝关节成骨细胞一氧化氮合酶表达和软骨下骨质重建的影响。方法取健康SD大鼠24只，体重（0．45±0．02）kg，随机分为实验组（A组）和对照组（B组），每组12只。实验组每周一次左膝关节腔内注射复方倍他米松0．02ml／kg，对照组每周一次左膝关节腔内注射生理盐水0．02ml／kg。4周时对两组大鼠的左侧膝关节股骨成骨细胞的诱生型一氧化氮合酶（inducible nitric oxide synthase，iNOS）和内皮细胞型一氧化氮合酶（endothelial nitric oxide synthase，eNOS）免疫组织化学研究，测定其平均灰度值和平均积分光密度，同时进行左胫骨软骨下骨质显微结构观察。结果实验组和对照组iNOS免疫组化的结果均为阴性，实验组eNOS的表达显著低于对照组（P〈0．01），实验组左胫骨的软骨下骨质旱骨质疏松特征。结论糖皮质激素对膝关节成骨细胞内皮细胞型一氧化氮合酶的表达和软骨下骨质生成有明显的抑制作用。     

15-脂氧酶-1基因表达调节研究进展

15-脂氧酶-1（15-LOX-1）是脂质过氧化物酶，分别以亚油酸、花生四烯酸为底物氧化生成13-羟基-十八碳二烯酸（13-HODE）、15-羟基-二十碳四烯酸（15-HETE）。15-LOX-1基因表达在转录、翻译、翻译后修饰各个水平都受到多种因素的调节，影响着15-LOX-1的表达量和酶催化活性。本文就15-LOX-1转录、翻译、翻译后修饰各个环节的调节因素作一简要介绍。     

中国Leber遗传性视神经病变G11696A突变两个家系分析

目的对两个中国Leber遗传性视神经病变(Leber’shereditary optic neuropathy,LHON)家系的临床和分子遗传学特征进行分析。方法眼科临床检查发现在这两个家系中只有先证者1人出现视力障碍,发病年龄分别为10岁和17岁。对这两个家系先证者使用24对有部分重叠的引物进行线粒体DNA(mitochondrial DNA,mtDNA)全序列扩增分析。结果没有发现mtDNAG11778A、G3460A和T14484C3个常见的突变位点,而发现了与LHON相关的ND4G11196A同质性突变位点的存在,在167名正常对照只发现1例G11696A突变。结论线粒体DNA全序列分析发现两个家系呈现独特的mtDNA多态性,都属于东亚单体型D4。不完全外显率和正常对照频率(1/167)表明G11696A突变本身不足以导致LHON的发生,说明其它因素在这两个LHON家系的表型表达中也起一定的作用。在这些家系mtDNA中缺乏影响重要功能突变位点的存在,排除了线粒体背景对LHON临床表型的影响。因此,核修饰基因、环境因素可能对两个中国G11696A突变家系的外显率和发病严重程度起促进作用。