Ginseng, the root of Panax ginseng, has been used as folk medicine in the treatment of various diseases for thousands of years in China.Ginsenoside Rb1(Rb1), one of the effective components of ginseng, has been reported to release nitric oxide and decrease intracellular free Ca2+ in cardiac myocytes, both of which play important roles in antihypertrophic effect.This study was to investigate the potential effect of Rb1 on right ventricular hypertrophy (RVH) induced by monocrotaline (MCT) and its possible influence on calcineurin (CaN) signal trasnsduction pathway.MCT-treated animals were administered with Rb1 (10 and 40 mg /kg) from day 1 to day 14 (preventive administration) or from day 15 to day 28 (therapeutic administration), or with vehicle as corresponding controls.After 2 weeks, significantly hypertrophic reactions, including RVH index and the expressions of atrial natriuretic peptide mRNA, appeared in right ventricle of all MCT-treated animals (p<0.05), which were significantly decreased with some improvements of myocardial pathomorphology in both Rb1 prevention- and therapy-groups(p<0.05).Similarly, MCT-treatment caused the high expressions of mRNA and/or proteins of CaN, NFAT3 and GATA4 from cardiocytes(p<0.05)and Rb1 could alleviate the expressions of these factors above (p<0.05).These results suggest that Rb1 treatment can inhibit the RVH induced by MCT, which may be involved in its inhibitory effects on CaN signal transduction pathway.

目的构建高效表达人过氧化物酶体增殖物激活受体δ（hPPARδ）的真核表达载体，为hPPARδ受体功能和基于hPPARδ受体靶点的药物筛选提供分子研究平台。方法 采用逆转录一聚合酶链式反应（RT-PCR），从HepG2细胞总RNA克隆hPPARδ全长基因，与经BamHI、Sa/I相同双酶切的pIRES2-EGFP载体连接，构建重组质粒phPPARδ-IRES2-EGFP，经酶切及基因测序鉴定重组质粒中hPPARδ基因的完整性和忠实性；荧光显微镜观察重组质粒转染的293细胞GFP报告基因表达强度，并对转染细胞hPPARδ的表达进行荧光定量PCR和免疫细胞化学检测。结果 经酶切和测序证实重组赝粒构建正确，并在转染的293细胞巾获得hPPAR6的高效表达。结论 成功构建phPPARδ-IRES2-EGFP重组质粒。

目的 构建可以高效表达人过氧化物酶体增殖物激活受体（αahPPARa）的真核细胞表达载体，为筛选作用于PPARa受体的药物提供分子平台。方法将HepG2细胞总RNA经逆转录一聚合酶链式反应（RT-PCR）克隆hPPARα全长基因，用BamHI、SalI双酶切后，与相同双酶切的pIRES2-EGFP载体连接，构建phPPARα-IRE52-EGFP重组质粒，用酶切及基因测序鉴定重组质粒中hPPARα綦因的完整性和可靠性；重组质粒转染293细胞，荧光显微镜观察EGFP报告基因表达强度，并对转染细胞的hPPARα表达进行荧光定量PCR及免疫细胞化学检测。结果 经酶切和测序证实重组质粒构建正确，并在转染的293细胞中获得hPPARα的高效表达。结论 成功构建重组质粒phPPARa-IRES2-EG-FP，为基于hPPARα受体靶点的药物筛选平台的建立提供了高效表达hPPARα的重组载体。

目的建立用于测定大鼠血清中喹啉酸含量的气质联用（gas chromatography- mass spectrometrv，CC-MS）技术。方法 动物分为对照组和炎症模型组。经内眦静脉取血，对采集的血清样品进行纯化和衍生化，采用GC-MS联用技术测定喹啉酸含量，以全扫描方式定性，选择性离子监测（selected ion monitoring，SIM）方式定量检测。结果 测定方法的专一性良好，检出限为50 nmol/L，喹啉酸浓度在0.05-20 ymol/L范围内线性关系良好。结论 本方法检测结果准确、T扰少，检定快速，适合需要多次少量取血的动物实验应用。喹啉酸的六氟异丙醇衍生物较为稳定，但血清不宜长时间冻存，宜尽怏测定。

目的：研究过氧化物酶体增殖物激活受体叫（PPAR7）激动剂罗格列酮对人乳腺癌细胞MDA-MB-231体外生长影响，以评价其在人乳腺癌治疗上的应用潜力．方法：噻唑蓝（ MTT）法检测罗格列酮对MDA-MB-231绌胞体外生长抑制作用；应用PPARα拮抗剂GW9662，分析罗格列酮对MDA-MB-231细胞增殖影响与PPARγ受体关系；流式细胞仪分析细胞周期，Annexin V-FITC和TUNEL法检测细胞凋亡．结果：罗格列酮T预下MDA-MB-231细胞Go/G，期比例上升，而S期比例下降，呈量一效关系抑制其生长，IC50=5.2ymol/L.PPARγ拮抗剂GW9662可部分逆转罗格列酮对MDA-MB-231细胞增殖抑制作用（P<0.05）；100 pdmol/L罗格列酮还可诱导MDA-MB-231细胞凋亡，TUNEL法检测的凋亡率（18.4±3.1）%与Annexin V-FITC法检测的结果一致[（16.6±2.7）%l（P>0.05）．结论：罗格列酮通过PPARγ介导，使MDA-MB-231细胞G．期阻滞而抑制其增殖，高浓度时还可诱导其发生凋亡，罗格列酮有望成为治疗乳腺癌的有效药物．

Surface molecules are important biomarkers for cell proliferation and differentiation and play important roles in cell function and cell interaction.Notch is a transmembrane receptor that regulates developmental processes and cell-fate decision.Histamine is used as an adjunct to immunotherapy in myelogenous leukemia, and regulates hematopoietic cell development.Thus, we investigated the effects of histamine on immunophenotype and Notch signaling in human HL-60 leukemia cells.Histamine(0.1-10lM)inhibited the colony-forming efficiency of HL-60 cells in a dose-dependent fashion and shifted the growth curve to the right.HL-60 cells were treated with histamine 0.1-1.0lM for 6days, and surface molecules were analyzed by flow cytometry. Histamine decreased CD49d positive cells by 74% while increasing CD31 positive cells by 53% as compared to controls.Histamine did not affect the expression of CD11b, CD14, CD34, CD44, CD54, CD49e, and CD62L.To examine Notch signaling in histamine-induced immunophenotype alterations in HL-60 cells, total RNA was isolated, purified, and subjected to real-time RT-PCR analysis. The expressions of Notch1, Notch4, the ligands Jagged1, Delta4, and the downstream hairy enhancer of split 1 gene (HES1) were not significantly altered by histamine.In summary, this study demonstrated that histamine inhibited HL-60 cell growth and regulated immunophenotypes of CD49d and CD31. These effects are not mediated through the Notch signaling.

The putative protective effects of meloxicam on the oxidative damage induced by aluminum overload in mice brain were investigated.The cerebral damage model in mice was established via intracerebroventricular(i.c.v.)microinjection of aluminum(5.0μg in 2.0μl), once a day for 5 days.Meloxicam, a selective inhibitor of cyclooxygenase-2(COX-2), was intragastrically(i.g)administered 30min before each aluminum administration, and continuously given for another 10days after the last aluminum administration.Behavioral changes including locomotor activity, passive avoidance, and spatial learning and memory ability were examined two weeks after the last administration of meloxicam.To determine the brain damage, we also measured pathological alterations in the cerebral tissue, malondialdehyde contents and expressions of Choline acetyltransferase (ChAT), amyloid precursor protein(APP)and amyloid beta.Furthermore, COX-2 proteins and COX-2 mRNA were examined to investigate the mechanism for underlying the effect of meloxicam.The impairment of learning and memory function was caused by aluminum overload.Consistent with the behavioral changes, neuronal death in the hippocampi, increased content of malondialdehyde, expressions of APP, amyloid beta and COX-2 proteins, as well as COX-2mRNA, and decreased expression of ChAT protein were detected in the aluminumoverload mice.Meloxicam significantly protected mice from the brain damage, and behavioral and biochemical changes above caused by aluminum overload. These experimental results indicate that there is a close relationship between over-expression of COX-2 and neuron damage induced by aluminum overload.It also suggests that selective inhibitors of COX-2 have potential values in clinical treatment for some other neuron damage-related diseases.

目的证明小檗碱抗肿瘤作用机制可能与信号转导过程的调控有关。方法采用细胞增殖抑制和Hoechst 33258染色凋亡实验比较小檗碱和黄连总碱对人结肠癌HCT116和SW480细胞的作用。利用Tcf-4报眚基因研究小檗碱对肿瘤细胞的信号转导影响。结果小檗碱在5-40 mg•L-l浓度范围内呈浓度依赖性和时间依赖性抑制人结肠癌HCT116和SW480细胞的增殖；小檗碱（20mgL-l）处理72 h后的HCT116和SW480细胞出现明显凋亡；相当于小檗碱浓度的黄连总碱有类似于小檗碱的作用。20-40 mg•L-l小檗碱和黄连总碱均能明显抑制β-catenin/Tcf介导的转录活性。结论黄连总碱的抗肿瘤作用可能与其主要成分小檗碱有关；其抗肿瘤作用机制至少与抑制Wnt/β-catenin信号通路有关。

目的 设想神经元缺氧损伤与自由基损伤线粒体有关，由此探讨淫羊藿苷的保护作用机制。方法 采用Fe2+/维生素C（VitC）为氧自由基生成系统建立氧自由基损伤线粒体的体外模型。观察淫羊藿苷对线粒体肿胀度、呼吸链复合体酶I~Ⅳ活性、丙二醛（MDA）含量的影响。结果 Fe2+ /VitC（lmmol•L-1/1 mmol•L-1，10 mmoL-1/10 mmol•L-1）可使线粒体的肿胀度和MDA含量显著增加，呼吸链复合体酶Ⅱ~Ⅳ活性不同程度下降。预先加入淫羊藿苷（0.03和0.1mg•L-1）能显著抑制线粒体肿胀，减少MDA含量，提高呼吸链复合体酶Ⅱ~Ⅳ的活性。结论 淫羊藿苷对氧自由基损伤酌大鼠脑线粒体呼吸链具有保护作用。