人血管内皮生长因子-C基因治疗淋巴水肿的实验研究

目的 观察人血管内皮生长因子-C(VEGF-C)基因治疗阻塞性淋巴水肿的疗效.方法 成年新西兰大白兔制成右后肢淋巴水肿模型;SD大鼠制成尾巴淋巴水肿模型.每种动物随机分2组,治疗组皮内注射人VEGF-C基因,对照组皮内注射生理盐水.测量治疗前后水肿部位体积变化;第3周取材,测VEGF-C mRNA及蛋白的表达水平;用淋巴管内皮细胞透明质酸受体-1(LYVE-1)抗体免疫组化染色观察淋巴管增生情况.结果 基因治疗后兔右后肢体积明显减少,其减少值分别为:治疗组(24.40±1.08)ml,对照组(5.80±1.92)ml,差异有统计学意义(P=0.0001);治疗组VEGF-C mRNA及蛋白明显表达,而对照组则不明显;治疗组淋巴管数量显著多于对照组(P=0.0004),且管腔较粗.结论 用VEGF-C基因治疗可诱导淋巴管生成,减轻或消除淋巴水肿.     

Integrated gene expression profiling analysis reveals SERPINA3, FCN3, FREM1, MNS1 as candidate biomarkers in heart failure and their correlation with immune infiltration

BackgroundThe purpose of this study was to identify possible diagnostic indicators for heart failure (HF) and to investigate the function of immune cell infiltration in this pathophysiology.MethodsHF datasets from the Gene Expression Metascape database were utilized. R software was used to the identify differentially-expressed genes (DEGs) and perform functional correlation analysis. Least absolute shrinkage and selection operator (LASSO) and Boruta algorithms elimination algorithms were then employed to screen and validate the HF diagnostic variables. Finally, Single-sample Gene Set Enrichment Analysis (ssGSEA) was utilized to assess immune cell infiltration in HF tissues, and the Spearman association between gene expression and immune cell concentration was investigated.ResultsA total of 239 DEGs were screened in this study. SERPINA3 (area under the curve, AUC =0.958), FCN3 (AUC =0.972), FREM1 (AUC =0.954), and MNS1 (AUC =0.948) were identified as diagnostic factors of HF. The gene set differentiation analysis (GSVA) (R package “GSVA”) results showed that the high expression of FREM1 and MNS1 genes was involved in bile acid, fatty acid, and heme metabolism, suggesting that the core gene affects the progression of HF by regulating metabolism. Meanwhile, the high expression of FCN3 and SERPINA3 was related to xenobiotic metabolism, inflammatory response, and adipogenesis.ConclusionsGiven the importance of immune cell infiltration in the genesis and progression of HF, SERPINA3, FCN3, FREM1, and MNS1 may be used as diagnostic variables for HF.     

人动脉粥样硬化斑块p53基因突变的初步报告

从甲醛固定的成人动脉粥样硬化斑块组织中提取ＤＮＡ，用ＰＣＲ扩增ｐ５３基因片段，将扩增的ｐ５３基因片段亚克隆至ＳＫ载体内，对扩增后的片段进行核苷酸序列分析。结果发现，动脉粥样硬化斑块中ｐ５３基因内第八外显子中第２７２位密码子的Ｔ变为Ｃ。免疫组化分析观察到斑块内有ｐ５３基因突变蛋白的表达。提示：ｐ５３基因突变可能与动脉粥样硬化的细胞增殖异常有关。     

Human Cytochrome P450 Family 4 Enzymes: Function,Genetic Variation and Regulation

The microsomal cytochrome P450 (CYP) family 4 monooxygenases are the major fatty acid ω-hydroxylases. These enzymes remove excess free fatty acids to prevent lipotoxicity, catabolize leukotrienes and prostanoids, and also produce bioactive metabolites from arachidonic acid ω-hydroxylation. In addition to endogenous substrates, recent evidence indicates that CYP4 monooxygenases can also metabolize xenobiotics, including therapeutic drugs. This review focuses on human CYP4 enzymes and updates current knowledge concerning catalytic activity profiles, genetic variation and regulation of expression. Comparative differences between the human and rodent CYP4 enzymes regarding catalytic function and conditional expression are also discussed.     

In vivo perivascular implantation of encapsulated packaging cells for prolonged retroviral gene transfer

Long-term benefits of coronary angioplasty remain limited by the treatmentinduced renarrowing of arteries, termed restenosis. One of the mechanisms leading to restenosis is the proliferation of smooth muscle cells. Therefore, proliferating cells of the injured arterial wall, which can be selectively transduced by retroviruses, are potential targets for gene therapy strategies. A direct single-dose therapeutic application of retroviral vectors for inhibition of cell proliferation is normally limited by too low transduction efficiencies. Encapsulated retrovirus-producing cells release viral vectors from microcapsules, and may enhance the transduction efficiency by prolonged infection. Primary and immortal murine and porcine cells and murine retrovirusproducing cells were encapsulated in cellulose sulphate. Cell viability was monitored by analysing cell metabolism. Safety, stability, transfer efficiency and extent of restenosis using capsules were determined in a porcine restenosis model for local gene therapy using morphometry, histology, in situ betagalactosidase assay and PCR. Encapsulation of cells did not impair cell viability. Capsules containing retrovirus-producing cells expressing the beta-galactosidase reporter gene were implanted into periarterial tissue or a pig model of restenosis. Three weeks following implantation, beta-galactosidase activity was detected in the pericapsular tissue with a transduction efficiency of ~ 1 in 500 cells. Adventitial implantation of vector-producing encapsulated cells for gene therapy may, therefore, facilitate successful targeting of proliferating vascular smooth muscle cells, and allow stable integration of therapeutic genes into surrounding cells. The encapsulation of vector-producing cells could represent a novel and feasible way to optimize local retroviral gene therapy.     

Genetics of inflammation and atopy

Multiple chromosomal regions and polymorphisms of several candidate genes have been linked to or associated with atopic diseases (hayfever, asthma, allergic eczema and rhinitis). In this mini-review, we present data demonstrating that the genetic regulation of the inflammatory response makes a major contribution to the risk of atopy. These data also suggest that the quantity (or quality) of the inflammation affects the priming phase of atopy, i.e., that induced by allergens or infectious agents in early childhood.     

河北汉族人群 CYP1A2 C163A基因多态性研究

目的：探讨河北汉族人群细胞色素P4501A2（cytochrome P4501A2,CYP1A2）C163A基因多态性的分布情况。方法应用聚合酶链式反应－限制性酶切片段长度多态性（PCR‐RFLP）方法对210例河北汉族正常人进行CYP1A2 C163A基因多态性分析。结果河北汉族人群CYP1A2 C163A基因A和C等位基因的频率为：68．3％、31．7％。AA、AC、CC基因型频率分别为48．1％、40．5％、11．4％。符合 Hardy‐Weinberg 遗传平衡定律（χ2＝1．22,P＞0．05）,具有代表性。结论河北汉族人群CYP1A2基因存在C163A位点多态性。     

腺病毒介导VEGF启动子驱动的TK及CD融合基因系统与碘油混合栓塞治疗兔肝癌的实验研究

目的 评价携带血管内皮生长因子(VEGF)启动子调控的TK及CD融合双自杀基因重组腺病毒系统(AdVEGF-CDglyTK)与超液化碘油混合栓塞兔肝癌后的治疗效果及安全性. 资料与方法 36只荷VX2瘤兔随机分4组,LP组(单纯超液化碘油栓塞组);LP AdVEGF-CDglyTK组(超液化碘油 AdVEGF-CDglyTK栓塞混合栓塞,介入术后腹腔注射GCV 5-FC治疗组);AdVEGF-CDglyTK组(单纯灌注AdVEGF-CDglyTK,介入术后腹腔注射GCV 5-FC治疗组);NS组(生理盐水治疗组).于术前及术后第10、15天行MRI检查观察肿瘤的体积,免疫组织化学法检测肿瘤组织VEGF的表达及微血管密度(MVD). 结果 4组兔术前肿瘤体积在统计学无显著差异(P>0.05);介入治疗后10天、15天的各组之间肿瘤增长率均有显著性差异(P<0.05),LP AdVEGF-CDglyTK肿瘤增长率最小.VEGF表达方面:LP组表达较其他三组高(P<0.05).MVD:LP AdVEGF-CDglyTK组较其他组低(P<0.05). 结论 在对兔肝癌的治疗中,与单纯碘油栓塞以及单纯灌注AdVEGF-CDglyTK相比,AdVEGF-CDglyTK与碘油混合肝动脉栓塞可以明显降低肿瘤的生长率,减少肿瘤组织VEGF的表达及减少肿瘤新生血管生成.