玉米苞叶对动脉粥样硬化家兔平滑肌细胞凋亡及p53和Fas蛋白表达的影响

目的：探讨玉米苞叶防治动脉粥样硬化（AS）的机制。方法：选用大耳白家兔，复制家兔AS模型，随机分为动脉粥样硬化模型组，玉米苞叶组和正常对照组，成模后给予玉米苞叶煎剂治疗，8w后处死家兔，采用流氏细胞术检测平滑肌细胞的凋亡率以及凋亡相关基因p53和Fas蛋白表达。结果：模型组血管平滑肌细胞的凋亡率明显高于对照组（P＜0．05），p53和Fas蛋白的表达增强（P＜0．05），主动脉壁肉眼观测出现典型斑块。玉米苞叶组平滑肌细胞的凋亡率明显低于模型组（P＜0．05），p53和Fas蛋白表达下调（P＜0．05）；主动脉斑块面积较模型组明显减小，结论：玉米苞叶通过调节p53和Fas蛋白表达而调节AS家兔平滑肌细胞凋亡。     

玉米苞叶降低动脉硬化家兔白细胞凋亡及CD44表达

目的　探讨玉米苞叶对动脉粥样硬化模型动物外周血白细胞凋亡和CD4 4表达的影响。方法　将家兔分为正常对照组、动脉硬化模型组、低剂量组 (造模 9w后给予低剂量玉米苞叶煎剂 )、高剂量组 (造模 9w后给予高剂量玉米苞叶煎剂 )。 1 7w后测血脂和血中白细胞凋亡率及CD4 4表达量 ,观察主动脉形态变化。结果　①模型组白细胞凋亡率 (6 58± 2 53) % ,CD4 4表达量 6 97± 1 41 ,均显著高于正常组 (P <0 0 1 ) ,②白细胞凋亡率和CD4 4表达量在低剂量组分别为 3 1 1± 1 50、4 76± 0 57;高剂量组为 3 74± 1 66、4 64± 0 42 ,均显著低于模型组 (P <0 0 1 )。结论　玉米苞叶煎剂可显著降低动脉硬化家兔白细胞凋亡率和CD4 4表达量 ,且这种作用不是通过降血脂实现的。     

玉米苞叶煎剂对动脉粥样硬化家兔血管外器官病变的形态学影响

目的 观察玉米苞叶煎剂对AS家兔血管外各器官病变的形态学影响.方法 选用大耳白家兔,雌雄各半,随机分为对照组、高脂组、玉米苞叶煎剂组,复制高脂食饵性家兔AS模型,观察玉米苞叶煎剂预防性用药后对家兔血脂及内脏各器官的病理形态学影响.结果 玉米苞叶煎剂组TC、TG较高脂组明显降低(P<0.01);高脂组各脏器(心、肝、脾、肺、肾)与对照组相比均有明显病理变化,玉米苞叶煎剂组各脏器病变较高脂组明显减轻.结论 玉米苞叶煎剂不但可以减轻AS家兔血管病变,且对心、肝、脾、肺、肾各脏器的高脂损伤同样有干预作用.     

玉米苞叶煎剂对高脂血症模型大鼠血管内皮及平滑肌增殖与凋亡的干预作用

目的 观察高脂血症大鼠主动脉内皮和平滑肌细胞增殖与凋亡对玉米苞叶煎剂作用的药效反应.方法 选用雄性SD大鼠,随机分为正常对照、病理模型、阳性对照及玉米苞叶煎剂组,复制食饵性高脂血症,玉米苞叶煎剂预防性给药,以吉非罗齐为阳性药对照.观察血脂及主动脉壁病理变化,流式细胞术定量检测内皮及平滑肌细胞增殖指数及凋亡率.结果 玉米苞叶煎剂组总胆固醇(TC)、甘油三酯(TG)、极低密度脂蛋白(VLDL)较模型组降低明显(均P＜0.05);与模型组相比,内皮凋亡率下降和平滑肌凋亡率升高均有显著性差异(均P＜0.05),内皮增殖指数轻度上升(P＞0.05),平滑肌增殖指数低于模型组(P＜0.05).结论 玉米苞叶煎剂可通过调节脂代解,阻断AS早期的内皮-平滑肌效应.     

益气活血中药对糖尿病动脉粥样硬化家兔血管平滑肌细胞凋亡及Bcl-2表达的影响

目的探讨益气活血中药对糖尿病动脉粥样硬化家兔血管平滑肌细胞凋亡及凋亡相关基因Bcl-2表达的影响。方法选用新西兰家兔,利用静脉注射胰岛β细胞毒剂四氧嘧啶,继以高糖高脂饲料复制家兔糖尿病动脉粥样硬化模型,随机分为模型组、小剂量中药治疗组、大剂量中药治疗组和正常对照组,实验12周后,处死家兔,观察主动脉内膜的病理形态学改变,采用流式细胞术检测凋亡率以及凋亡相关基因Bcl-2表达。结果模型组与正常对照组比较,动脉内膜出现典型的斑块,动脉管腔极度狭窄,平滑肌细胞凋亡率明显增高(13.89%±1.53%比2.36%±0.76%,P<0.01),Bcl-2表达降低(22.04±1.38比30.48±0.68,P<0.01)。中药治疗组与模型组比较,主动脉斑块缩小,动脉管腔狭窄减轻,平滑肌细胞凋亡率明显降低(P<0.01),Bcl-2表达上调(P<0.05),且大剂量中药治疗组优于小剂量中药治疗组。结论益气活血中药拮抗糖尿病动脉粥样硬化形成的机制可能与上调Bcl-2蛋白的表达,抑制血管平滑肌细胞凋亡有关。     

玉米苞叶煎剂对动脉粥样硬化家兔血清内皮素、前列环素及病理形态学影响

为研究玉米苞叶对动脉粥样硬化的治疗作用 ,通过建立高脂食饵性兔动脉粥样硬化模型 ,观察了玉米苞叶煎剂治疗用药后对血清内皮素、6 酮 前列腺素及主动脉粥样硬化病理形态学的影响。结果发现 ,模型组血清内皮素和 6 酮 前列腺素较对照组均明显降低 ,每日饲饮含硅量 2mg (kg·d)玉米苞叶煎剂 10 0mL 只 (低剂量组 )或含硅量 4mg (kg·d)玉米苞叶煎剂 10 0mL 只 (高剂量组 )。于实验 17周末发现 ,血清内皮素较模型组有所上升 ,6 酮 前列腺素F1а较模型组明显升高 (模型组为 6 6± 2 6 ,低剂量组为 12 2 .4± 2 .4 ,高剂量组为 117± 6 4 ,P <0 .0 5 ) ,内皮素与前列环素比值也明显降低 (模型组为 7.0± 1.2 ,低剂量组为 5 .1± 1.4 ,高剂量组为 5 .7± 1.3,P <0 .0 5 ) ,主动脉内膜粥样斑块病变程度和面积较模型组明显减轻和缩小。结果提示 ,玉米苞叶煎剂具有保护动脉内膜损伤 ,调节内皮细胞血管活性物质分泌 ,治疗家兔动脉粥样硬化作用     

白皮杉醇对过氧化氢诱导的血管平滑肌细胞凋亡的影响与机制

目的:研究白皮杉醇(PCT)对过氧化氢(H2O2)诱导的血管平滑肌细胞(VSMCs)凋亡的影响以及所涉及的相关分子机制。方法:组织贴块法培养SD大鼠VSMCs并进行不同分组与处理;MTT法检测各组VSMCs的存活率;LDH、超氧化物歧化酶(SOD)试剂盒测定各组VSMCs中LDH与SOD活力;Annexin V-PI染色检测各组VSMCs凋亡情况,Hoechst 33258染色观察各组VSMCs细胞核形态变化;Western blot检测各组VSMCs中凋亡相关蛋白(Bcl-2、Bax、caspase-3)及PI3K/AKT/eNOS通路蛋白表达水平。结果:不同浓度与时间过氧化氢(H2O2)处理VSMCs后使其存活率降低,而不同浓度PCT预处理可提高细胞存活率,差异有统计学意义(P<0.05、P<0.01或P<0.001)。与对照组相比,H2O2组VSMCs中LDH活力升高而SOD活力下降,细胞凋亡率增加,细胞中Bax、caspase-3蛋白表达升高而Bcl-2蛋白表达下降,p-PI3K、p-Akt、p-eNOS蛋白表达下降,且均呈剂量依赖关系;与H2O2组相比,PCT预处理组VSMCs LDH活力下降而SOD活力升高,细胞凋亡率降低,细胞中Bax、caspase-3下降,Bcl-2蛋白表达升高,p-PI3K、p-Akt、p-eNOS的蛋白表达升高,且均呈剂量依赖关系,差异有统计学意义(P<0.05或P<0.01)。结论:PCT对H2O2引起的VSMCs损伤有保护作用,其机制可能是抑制LDH活力、增强SOD活力以及增强PI3K/AKT/eNOS信号通路进而阻碍VSMCs凋亡的发生。     

卡维地洛对自发性高血压大鼠心肌细胞凋亡及相关因素的影响

目的探讨卡维地洛对自发性高血压大鼠(SHR)心肌细胞凋亡、心肌Bcl-2、p53蛋白表达、血浆超氧化物歧化酶(SOD)、丙二醛(MDA)及血管紧张素Ⅱ(AngⅡ)水平的影响.方法6周龄雄性SHR12只,随机分为2组(n=6)对照组普通饲料喂养+NS 5 mL/d灌胃;卡维地洛组卡维地洛(Car)20 mg/(kg·d)+NS 5 mL溶解灌胃,其余同对照组,共8周.随后给予2%戊巴比妥钠40 mg/kg IP麻醉,剖腹取下腔静脉血3 mL,分3等分于试管中分离血浆保存,待测AngⅡ、SOD与MDA.继之开胸取心脏,取左心室游离壁心肌于10%中性福尔马林中固定24 h,继之脱水,石蜡包埋及切片,并采用TUNEL法、免疫组化法分别检测心肌细胞凋亡率,Bcl-2及p53蛋白表达率.结果与对照组比较,Car组血浆MDA、AngⅡ、心肌细胞凋亡率及心肌细胞p53蛋白表达率均明显降低,而血浆SOD、心肌细胞Bcl-2蛋白表达率则明显增高(P＜0.01).且两组中MDA、AngⅡ及p53蛋白表达率与心肌细胞凋亡率均分别呈正相关;而SOD、Bcl-2蛋白表达率与心肌细胞凋亡率呈负相关(P＜0.05).结论Car具有明显抑制SHR心肌细胞凋亡,p53蛋白表达和抗氧化作用,同时还能明显改善Bcl-2蛋白表达的效应,AngⅡ、p53蛋白表达、Bcl-2蛋白表达和脂质过氧化反应均可能参与了心肌细胞凋亡过程.     

麝香通心滴丸对稳定动脉粥样硬化斑块的机制研究

目的观察麝香通心滴丸稳定动脉粥样硬化(AS)斑块的作用机制。方法采用高脂饮食伴主动脉内膜球囊损伤复制家兔动脉粥样硬化斑块模型,药物(斑点蝰蛇毒、组胺)触发斑块破裂,观察麝香通心滴丸稳定易损斑块的作用机制。生化方法检测AS家兔的血脂及氧化应激水平;ELISA及实时荧光定量PCR(Real-Time RT-PCR)法检测麝香通心滴丸对AS家兔基质金属蛋白酶-2(MMP-2)、TIMP2以及相关炎症介质的蛋白含量和mRNA表达水平的影响;应用TUNEL法检测麝香通心滴丸对AS家兔斑块内细胞凋亡水平的影响。结论麝香通心滴丸稳定易损斑块作用的机制有,减轻AS氧化应激损伤,降低脂质过氧化;降低MMP2含量,升高TIMP2含量,减少AS斑块内的基质降解,调节基质代谢;减少斑块内巨噬细胞的数量,降低炎性因子的表达,减轻斑块内的炎症反应;减少斑块内细胞凋亡增加斑块稳定性。     

丹参对粥样斑块Bax、Bcl-2和MCP-1、IL-6含量的影响

目的研究丹参抑制动脉粥样硬化形成的分子机制。方法建立动脉粥样硬化家兔模型,比较对照组、AS组及丹参组动脉粥样斑块形成的变化;免疫组化结合RT-PCR法分析Bcl-2、Bax及MCP-1、IL-6在粥样硬化斑块中的表达。结果丹参组血脂水平和动脉粥样斑块面积低于AS组(P<0.01);免疫组化显示,AS组Bcl-2、Bax及MCP-1、IL-6阳性细胞表达与正常组有显著差异。而丹参的干预可以调节Bax/Bcl-2阳性细胞表达率,同时下调MCP-1蛋白表达。结论丹参抑制动脉粥样硬化斑块的形成;调节Bcl-2/Bax表达及炎症机制可能是抗动脉粥样硬化的机制之一。     

妊娠时间与肺结核复发的相关性研究及诊治对策

目的 分析肺结核史患者妊娠时间和肺结核复发间相关性.方法 选取我院收治的有肺结核史的妊娠妇女576例作为研究对象,对其妊娠前肺结核治疗、治愈后妊娠时间、妊娠后复发肺结核等进行分析,总结有肺结核史育龄女性的妊娠时间和肺结核复发之间的关系.结果 肺结核治愈后不同时间段妊娠者的结核复发率比较,差异具有显著性（P＜0.05）,停药后间隔时间越久妊娠,肺结核复发的几率越小.结论 加强孕期痰菌检查,及早发现复发肺结核,提高母婴安全.     

我国骨关节结核诊治的回顾与展望

骨关节结核是危害人们健康的严重感染性疾病,近95％由他处结核病继发而来.罹患骨关节结核疾病后几乎均将致残,严重影响人们的健康、工作和生活.建国以来在党和国家的关心和支持下,骨关节结核的诊治水平取得了长足进步.时至今日,由于多种原因,学科发展和被重视程度受到一定的制约,同整个医疗行业的发展不相适应.回顾过去,展望未来,我们需要重新审视骨关节结核的诊治方法,努力推进骨关节结核诊疗技术的科学发展.     

Effect of phosphorylation of MAPK and Stat3 and expression of c-fos and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44~(MAPK), p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44~(MAPK), p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44~(MAPK) and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between P42/44~(MAPK) and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Raf/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44~(MAPK), c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

Overweight and Obesity and Progression of ADPKD

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Replication and Inhibitors of Enteroviruses and Parechoviruses

The Enterovirus (EV) and Parechovirus genera of the picornavirus family include many important human pathogens, including poliovirus, rhinovirus, EV-A71, EV-D68, and human parechoviruses (HPeV). They cause a wide variety of diseases, ranging from a simple common cold to life-threatening diseases such as encephalitis and myocarditis. At the moment, no antiviral therapy is available against these viruses and it is not feasible to develop vaccines against all EVs and HPeVs due to the great number of serotypes. Therefore, a lot of effort is being invested in the development of antiviral drugs. Both viral proteins and host proteins essential for virus replication can be used as targets for virus inhibitors. As such, a good understanding of the complex process of virus replication is pivotal in the design of antiviral strategies goes hand in hand with a good understanding of the complex process of virus replication. In this review, we will give an overview of the current state of knowledge of EV and HPeV replication and how this can be inhibited by small-molecule inhibitors.     

Effect of phosphorylation of MAPK and Stat3 and expression of c-fas and c-jun proteins on hepatocarcinogenesis and their clinical significance

AIM To study the effect of phosphorylation ofMAPK and Stat3 and the expression of c-fos andc-jun proteins on hepatocellular carcinogenesisand their clinical significance.METHODS SP immunohistochemistry was usedto detect the expression of p42/44MAPK, p-Stat3,c-fos and c-jun proteins in 55 hepatocellularcarcinomas (HCC) and their surrounding livertissues.RESULTS The positive rates and expressionlevels of p42/44MAPK, p-Stat3, c-fos and c-junproteins in HCCs were significantly higher thanthose in pericarcinomatous liver tissues (PCLT).A positive correlation was observed between theexpression of p42/44MAPK and c-fos proteins, andbetween p-Stat3 and c-jun, but there was nosignificant correlation between p42/44MAPK and p-Stat3 in HCCs and their surrounding livertissues.CONCLUSION The abnormalities of Ras/Rat/MAPK and JAKs/ Stat3 cascade reaction maycontribute to malignant transformation ofhepatocytes. Hepatocytes which are positive forp42/ 44MAPK, c-fos or c-jun proteins may bepotential malignant pre-cancerous cells.Activation of MAPK and Stat3 proteins may be anearly event in hepatocellular carcinogenesis.     

心电图、心电向量图与冠状动脉造影结果对比分析

目的:通过分析心电图(Electrocardiogram,ECG)和心电向量图(Vectorcardiogram,VCG)的改变与冠脉造影（CAG）结果进行对比,探讨ECG、VCG在冠状动脉病变中的诊断价值。方法: 选择2008年1月~2009年12月临床拟诊断为冠心病患者108例,行常规ECG、VCG检查,并于1周内进行CAG,对检查结果依据各自的诊断标准进行判定,以CAG为标准诊断法,利用四格表法,计算相关评价真实性的指标并进行比较。结果: ①VCG检测的灵敏度、特异度、准确度显著高于ECG(P<0.05,P<0．01)。②ECG、VCG阳性率与冠脉病变支数组间比较:在单支病变、双支病变中,VCG阳性率明显高于ECG(P<0.05),左主干或三支病变无统计学意义;组内比较:ECG组左主干或三支病变组较单支病变、双支病变阳性率高(P<0.05,P<0.01);VCG组左主干或三支病变组较单支病变阳性率高(P<0.05);与双支病变阳性率比较无统计学意义;③ECG、VCG阳性率与冠脉病变程度组间比较:冠脉病变狭窄50%～69%的VCG阳性率明显高于ECG (P<0.05),其他两组阳性率比较无统计学意义;组内比较:ECG组冠脉病变狭窄≥90%较50%～69%、70%～89%的阳性率高(P<0.05,P<0.01); VCG组狭窄≥90%较50%～69%阳性率高（P<0.01),其他无统计学意义。结论: VCG对冠心病检测价值显著高于ECG。     

Detection and characterization of the product of hydroethidine and intracellular superoxide by HPLC and limitations of fluorescence

Here we report the structural characterization of the product formed from the reaction between hydroethidine (HE) and superoxide (O(2)(.-)). By using mass spectral and NMR techniques, the chemical structure of this product was determined as 2-hydroxyethidium (2-OH-E(+)). By using an authentic standard, we developed an HPLC approach to detect and quantitate the reaction product of HE and O(2)(.-) formed in bovine aortic endothelial cells after treatment with menadione or antimycin A to induce intracellular reactive oxygen species. Concomitantly, we used a spin trap, 5-tert-butoxycarbonyl-5-methyl-1-pyrroline N-oxide (BMPO), to detect and identify the structure of reactive oxygen species formed. BMPO trapped the O(2)(.-) that formed extracellularly and was detected as the BMPO-OH adduct during use of the EPR technique. BMPO, being cell-permeable, inhibited the intracellular formation of 2-OH-E(+). However, the intracellular BMPO spin adduct was not detected. The definitive characterization of the reaction product of O(2)(.-) with HE described here forms the basis of an unambiguous assay for intracellular detection and quantitation of O(2)(.-). Analysis of the fluorescence characteristics of ethidium (E(+)) and 2-OH-E(+) strongly suggests that the currently available fluorescence methodology is not suitable for quantitating intracellular O(2)(.-). We conclude that the HPLC/fluorescence assay using HE as a probe is more suitable [corrected] for detecting intracellular O(2)(.-).     

Effects of sex and time of day on metabolism and excretion of corticosterone in urine and feces of mice

Non-invasive techniques to monitor stress hormones in small animals like mice offer several advantages and are highly demanded in laboratory as well as in field research. Since knowledge about the species-specific metabolism and excretion of glucocorticoids is essential to develop such a technique, we conducted radiometabolism studies in mice (Mus musculus f. domesticus, strain C57BL/6J). Each mouse was injected intraperitoneally with 740 kBq of 3H-labelled corticosterone and all voided urine and fecal samples were collected for five days. In a first experiment 16 animals (eight of each sex) received the injection at 9 a.m., while eight mice (four of each sex) were injected at 9 p.m. in a second experiment. In both experiments radioactive metabolites were recovered predominantly in the feces, although males excreted significantly higher proportions via the feces (about 73%) than females (about 53%). Peak radioactivity in the urine was detected within about 2h after injection, while in the feces peak concentrations were observed later (depending on the time of injection: about 10h postinjection in experiment 1 and about 4h postinjection in experiment 2, thus proving an effect of the time of day). The number and relative abundance of fecal [3H]corticosterone metabolites was determined by high performance liquid chromatography (HPLC). The HPLC separations revealed that corticosterone was extensively metabolized mainly to more polar substances. Regarding the types of metabolites formed, significant differences were found between males and females, but not between the experiments. Additionally, the immunoreactivity of these metabolites was assessed by screening the HPLC fractions with four enzyme immunoassays (EIA). However, only a newly established EIA for 5alpha-pregnane-3beta,11beta,21-triol-20-one (measuring corticosterone metabolites with a 5alpha-3beta,11beta-diol structure) detected several peaks of radioactive metabolites with high intensity in both sexes, while the other EIAs showed only minor immunoreactivity. Thus, our study for the first time provides substantial information about metabolism and excretion of corticosterone in urine and feces of mice and is the first demonstrating a significant impact of the animals' sex and the time of day. Based on these data it should be possible to monitor adrenocortical activity non-invasively in this species by measuring fecal corticosterone metabolites with the newly developed EIA. Since mice are extensively used in research world-wide, this could open new perspectives in various fields from ecology to behavioral endocrinology.     

大鼠骨髓间充质干细胞的分离培养和外源基因的导入

目的探讨绿色荧光蛋白基因转染骨髓间质干细胞的可行性。方法采用F icoll-PaqueTMP lus淋巴细胞分离液,根据细胞密度梯度原理,分离大鼠骨髓间充质干细胞(rM SC s)并进行体外原代培养和传代扩增,倒置相差显微镜观察细胞生长情况,免疫细胞化学法对其初步鉴定。流式细胞仪分析转染效率。结果原代和传代培养的细胞呈现梭形外观,具有较强的生长增殖能力;细胞均一表达CD44、CD54、CD106、CD29抗原。电穿孔法转染rM SC s转染率为32.8%±3%。结论采用比重为1.077 g/L的F icoll-PaqueTMP lus能分离获得大鼠骨髓间充质干细胞,经原代培养和传代培养能够迅速扩增。电穿孔法具有较高的介导外源基因表达于rM SC s的效率。