大黄素对肾小球系膜细胞c—myc原癌基因表达的影响

观察大黄素对肾小素系膜细胞ｃ－ｍｙｃｍＲＮＡ表达的影响，探讨大黄素抑制ＭＣ生长的分子机理。网筛法分离大鼠肾小球，培养肾小球ＭＣ。ＡＧＰＣ一步法提取细胞总ＲＮＡ，ｃ－ｍｙｃｍＲＮＡ水平用斑点杂交法测定，以显著斑点最大的ＲＮＡ稀释度表示ｍＲＮＡ水平。     

蝮蛇抗栓酶对兔血管损伤后血管平滑肌细胞C—myc原癌基因表达的影响

目的：探讨蜊蛇抗栓酶抑制血管再狭窄的作用机制。对临床应用提供理论依据。为再狭窄的防治寻找治疗药物。方法：建立兔髂动脉再狭窄模型。用自行设计、全盛的C-myccDNA探针和原位杂交技术观察血管内膜C-mycmRNA的表达与内膜血管平滑细胞（cscularsmooth muscle cells,vSMCs)增殖和内膜形成的关系。结果：C-mycmRNA表达于动脉受损后1周即达高峰并分布于增生的内膜全层,平均阳性细胞个数为29.23.min^-2,蜊蛇抗栓酶1周时对C-mycmRNA的表达有明显的抑制作用,平均阳性细胞个数为10.73.mm^-2。结论：抑制C-myc基因的表达可能减少或阻止内明显病变的形成,有利于血管再狭窄的防治,研究显示蜊蛇抗栓酶对防治再狭窄有效。     

c—myc基因在食管异型增生组织和鳞癌中的表达及其意义

本研究通过单克隆抗体免疫组化LSAB法检测c-myc基因在正常食管粘膜(EM)、食管异型增生(EED)和食管鳞癌(SCCE)中表达的变化,目的是了解c-myc基因在食管癌发生和发展中的作用,及对临床的意义。 材料与方法 一、材料和试剂 1.选择35例食管癌术后标本,取EM10块(远癌5cra以外)、EED和SCCE各35块(分别取自癌旁0.5～1.0cm和癌中心)。标本用10％福尔马林固定和石蜡包埋,并记录年龄、发生部位、原发瘤大小、淋巴结状态、组织分化和PT参数,同时按1987年国际食管癌分期标准进行TNM分期。2.c-myc癌基因蛋白鼠抗人单抗NM-0409购于美国Novacastra(胞浆和核旁染色),LSAB试剂盒为美国Zymed公司生产。     

普罗布考预防经皮冠状动脉球囊成形术和经皮冠状动脉介入治疗术后再狭窄的研究进展

普罗布考（丙丁酚,probucol）是1977年首先在美国上市的一种降脂药,后在英、法、德、日等十几个国家相继上市,国内直在最近几年才开始使用,具有抗术后冠状再狭窄、调脂和抗氧化等作用。动物实验表明,普罗布考可减低动脉壁破损后内膜增生。在临床试验中,MVP（Multivitam ins and ProbucolTrial）试验显示,     

K—ras和C—myc在肺癌中表达的意义

目的：探讨原发性肺癌中K—ras和c—myc的表达与肺癌发生发展之间的关系。方法：应用免疫组化法测定53例肺癌标本及19例正常人的肺组织中K—ras和c-myc的表达。结果：肺癌细胞中K—ras和c—myc的阳性表达率明显高于正常人肺组P〈0．01）;K-ras和c—myc在不同病理类型（确切p=1）,和不同大小的肺癌组（P〉0．05）之间的差异无显著性;在三组不同分化程度的肺癌组间的阳性表达率差异有显著性（P〈0.05）;K—ras和c—myc在淋巴结有转移组的阳性表达率明显高于无转移组,差别有显著意义（P〈0．05）。结论：K—ras和c—myc蛋白的过度表达参与了人肺细胞癌的发生发展过程。     

小檗碱对局灶性脑缺血大鼠原癌基因c—fos表达的影响

目的：研究大鼠局灶性脑缺血过程中c-fos mRNA表达的动态变化，并观察小檗碱（Ber）对其作用。方法：运用斑点杂交技术，观察Ber对局灶性脑缺血再灌注大鼠大脑皮质及海马组织c-fos mRNA表达的影响。结果：大鼠局灶性脑缺血2h再灌注0．5－4h，脑皮质及海马组织c-fos mRNA出现一过性高表达，2h达高峰，分别为对照组的4．5和4．7倍。Ber 20mg/(kg·d)ip显抑制大鼠脑缺血诱导的c-fos高表达，降低病灶侧海马和皮层组织水、钙含量。结论：脑缺血过程中c-fos原癌基因呈现一过性高表达，Ber可降低c-fos mRNA水平，该作用可能是其抗脑缺血机理之一。     

普罗布考预防经皮冠状动脉介入治疗术后造影剂肾病的效果分析

目的 探讨普罗布考对经皮冠状动脉介入治疗(PCI)术后造影剂肾病(CIN)的预防作用。 方法 选取96例冠心病且均需行PCI术患者为研究对象,按随机数字表法将其分为对照组48例和观察组48例。对照组患者予常规治疗,观察组患者在常规治疗基础上加用普罗布考治疗。比较两组患者PCI术前和术后24 h、48 h、72 h时血清肌酐(Scr)、胱抑素C(CysC)、尿素氮(Bun)、肾小球滤过率(eGFR))以及C-反应蛋白(CRP)、白细胞介素6(IL-6)和丙二醛(MAD)水平,观察两组患者在治疗过程中的不良反应。 结果 两组术前Scr、CysC、Bun、eGFR差异均无统计学意义( t＝0.149, P＝0.882; t＝1.75, P＝0.243; t＝0.133, P＝0.894; t＝0.019, P＝0.985);两组术后Scr、CysC、Bun、eGFR差异均有统计学意义( t＝3.242, P＝0.002; t＝9.532, P<0.001; t＝9.073, P<0.001; t＝2.896, P<0.001),且Scr、CysC、Bun、eGFR水平均在术后24 h逐渐升高,至48 h达顶峰,72 h呈下降趋势,观察组升高及下降幅度均明显大于对照组。两组术前CRP、IL-6和MAD差异均无统计学意义( t＝0.321, P＝0.749; t＝0.014, P＝0.989; t＝0.188, P＝0.850),术后两组CRP、IL-6和MAD水平差异均有统计学意义( t＝8.495, P＝0.002; t＝7.532, P<0.00; t＝16.216, P<0.001),且其CRP、IL-6和MAD水平均在术后24 h逐渐上升,48 h达最高值,72 h呈下降趋势,且上升及下降趋势观察组均明显大于对照组。两组患者治疗过程中均无明显不良反应发生。 结论 普罗布考除对冠心病患者有调脂和抗动脉粥样硬化作用外,对行PCI术后的CIN有着一定预防效果,可作为临床上预防治疗CIN的优选药物之一。     

川芎嗪对球囊损伤后大鼠血管新生内膜增生的影响

目的探讨川芎嗪对大鼠颈总动脉球囊损伤后新生内膜增生的影响及其机制。方法 SD雄性大鼠32只,随机分为假手术组、模型组、川芎嗪高剂量组(60mg·kg~(-1)·d~(-1))和低剂量组(30mg·kg~(-1)·d~(-1))。用球囊导管损伤术建立大鼠颈总动脉内膜增生模型,测量给药14d后颈总动脉内膜面积(NIA)及内膜面积/中膜面积(NIA/MA),免疫组化法检测该动脉增殖细胞核抗原(PCNA)的表达,real-time RT-PCR法测定内皮型一氧化氮合酶(Nos3)和原癌基因c-Myc的mRNA表达,并检测血浆中超氧化物歧化酶(SOD)活性、丙二醛(MDA)以及环磷酸腺苷(cAMP)和环磷酸鸟苷(cGMP)的含量。结果川芎嗪高、低剂量组均能明显减少球囊损伤后大鼠颈总动脉的NIA及NIA/MA(P<0.05),并使PCNA蛋白质表达降低(P<0.01);显著升高球囊损伤大鼠血浆SOD活性和cAMP及cGMP的含量,而降低血浆MDA水平(均P<0.01);川芎嗪高剂量组还能上调Nos3 mRNA表达,而使c-Myc mRNA的表达降低(均P<0.05)。结论川芎嗪能抑制球囊损伤所致血管内膜异常增生,该作用可能与其抗氧化和促进环核苷酸生成有关。     

雷帕霉素抑制增生内膜中基质细胞衍生因子-1的表达

目的：进一步探讨雷帕霉素抗血管内再狭窄的分子机制。方法：球囊拉伤大鼠颈总动脉建立血管内膜增生的动物模型,口服雷帕霉素（25mg/kg）,4周后处死动物。HE染色观测血管病变,免疫组化检测增生内膜中基质细胞衍生因子-1（SDF-1）的表达,小室迁移系统观察SDF-1对平滑肌细胞迁移的影响。结果：球囊拉伤明显诱导血管内膜增生,增生内膜中有明显的SDF-1的表达,雷帕霉素能显著下调增生内膜中SDF-1的表达,而SDF-1浓度依赖性地诱导平滑肌细胞的迁移。结论：雷帕霉素可能通过下调SDF-1的表达从而抑制血管再狭窄。     

丙丁酚对高脂血症小鼠表达C反应蛋白的影响

目的观察丙丁酚对高脂高胆固醇饲养的高脂血症小鼠体内C反应蛋白（CRP）活性及表达的影响，探讨丙丁酚的作用机制。方法以高脂高胆周醇饲料饲喂小鼠，制作高脂血症模型。HE染色观察小鼠肝脏病变情况；采用相关检测试剂盒测定小鼠血脂水平、血浆中CRP和谷胱甘肽S转移酶的活性；采用逆转录聚合酶链式反应（RT—PCR）检测小鼠肝脏CRP mRNA的表达。结果高脂高胆固醇饮食显著增加C57BL／6J和apoE^./.小鼠血脂水平（P〈0．05），而丙丁酚则可降低高脂血症小鼠血脂水平（P〈0．05）。高脂组小鼠肝脏结构比较紊乱，脂肪空泡明显。与高脂组比较，经丙丁酚处理后，小鼠肝脏的脂肪空泡明显减少，并且其血浆中CRP的水平也下降（P〈0．05）。结论丙丁酚可以减轻高脂高胆固醇饲料诱导的高脂血症小鼠的CRP的水平，对肝脏产生抗炎性的保护作用。     

Effect of nilvadipine on balloon catheterization-induced intimal thickening of the coronary artery in miniature pigs.

The effect of nilvadipine on balloon catheterization-induced intimal thickening of the coronary artery was examined in miniature pigs. A diffuse intimal thickening was observed in vehicle-treated controls 6 weeks after balloon catheterization. The histologic features of the intimal thickening resembled those of early atherosclerotic lesions and restenosis after percutaneous transluminal coronary angioplasty (PTCA). Nilvadipine given in a daily dose of 10 mg/kg (subcutaneously, s.c.) for 6 weeks significantly inhibited intimal thickening. The ratio of the cross-sectional area of the intima to the cross-sectional area of the media was significantly reduced by 62% in nilvadipine-treated animals. These results suggest that nilvadipine may prevent or exert beneficial effects on coronary arterial injuries such as atherosclerosis and restenosis after PTCA.     

碘帕醇对糖尿病患者肾脏功能的影响

目的 探讨不同剂量碘帕醇对糖尿病患者肾功能的影响.方法 入选148例行冠状动脉造影或经皮冠状动脉介入(PCI)治疗的糖尿病患者,根据对比剂的使用量,分为低剂量组(＜100 ml),中剂量组(100～200 ml)和高剂量组(＞200 ml).术前及术后72 h检测患者血清肌酐浓度;分析不同剂量对比剂对血清肌酐水平、估测肌酐清除率(eGFR)的影响及年龄、性别和eGFR、低密度脂蛋白、血压水平等因素对对比剂肾病(CIN)发生率的影响.结果 术后各组间的肌酐浓度、eGFR差异无统计学意义(P＞0.05);术后肌酐水平在中、高剂量组明显高于术前[分别(96.4±21.8)μmol/L比(87.5±21.8)μmol/L,(100.8±23.9)μmol/L比(88.1±19.9)μmol/L,均P＜0.05],各组eGFR同术前相比差异无统计学意义(P＞0.05).低eGFR及女性患者CIN 发生率明显高于eGFR正常和男性患者(均P＜0.05).结论 在特定的糖尿病患者中使用较大剂量的碘帕醇是安全的.

Abstract：

Objective To determine the effects of the volume of contrast media and risk factors on kidney function in diabetic patients undergoing elective cardiac catheterization or percutaneous coronary intervention (PCI). Methods Totally 148 consecutive diabetic patients undergoing elective cardiac catheterization or PCI were divided into three groups; low dose (＜100 ml), middle dose (100-200 ml) and high dose group (＞200 ml). Serum creatinine and estimated glomerular filtration rate(eGFR) at baseline and after 72 hours of the procedure were measured. The incidence of CIN and the effect of variables on rate of CIN were evaluated. Results There were no significant differences on serum creatinine level and eGFR among groups after cardiac catheterization or PCI. Compared with the preoperative, the levels of serum creatinine in middle dose and high dose group were higher after the procedure [(96.4±21.8)μmol/L vs (87. 5 ±21. 8)μmol/L, (100. 8 ±23. 9) μmol/L vs (88.1 ± 19.9) μmol/L, P＜ 0.05]. The increase of serum creatinine in high dose group were significantly higher than in low dose and middle dose groups. There was no significant difference on eGFR and on the incidence of CIN among groups after cardiac catheterization or PCI. CIN developed more frequently after PCI in female or lower eGFR patients. Conclusion It is safe for some diabetic patients to use large dose of contrast media.     

Ultrasound microbubble-carried PNA targeting to c-myc mRNA inhibits the proliferation of rabbit iliac arterious smooth muscle cells and intimal hyperplasia

Objective: To elucidate the transfected effect of albumin ultrasound microbubbles carrying peptide nucleic acids (PNAs) against c-myc gene to the vascular walls and their effect on the intimal proliferation induced by vascular denudation.

Methods: A rabbit iliac artery intimal proliferation model was constructed and PNA against c-myc mRNA was designed and synthesized and was added to albumin solution before ultrasound microbubbles were prepared and encapsulated in matrix of albumin. The ultrasound microbubbles carrying PNA were transfected to intima under ultrasound exposure. The transfected effect was identified by a histochemical method and the expression of c-myc was detected by in situ hybridization. The proliferation of intimal smooth muscle cells was estimated by the expression of proliferative cell nuclear antigen (PCNA) of them. The intimal area and thickness were judged morphologically for intimal hyperplasia.

Results: The ultrasound microbubbles with PNA were successfully prepared and c-myc PNA was transfected to vascular intimal cells. The expression of c-myc and PCNA by intimal vascular smooth muscle cells (vSMCs) was inhibited significantly and the intimal thickness and area were reduced remarkably.

Conclusion: Transfection of c-myc PNA could inhibit proliferartion of vSMCs and intima in the rabbit iliac artery intimal proliferation model and the targeted transfection of albumin ultrasound microbubbles carrying PNA offers a feasible way to facilitate its access to specific cells in vivo and produce bioavailability.     

Neutrophil accumulation promotes intimal hyperplasia after photochemically induced arterial injury in mice

The role of leukocytes in the pathogenesis of coronary arterial disease has become a focus for clinical research. The aim of this study was to determine whether neutrophil accumulation would participate in the development of intimal hyperplasia after endothelial injury in mice, and whether d-myo-inositol hexakisphosphate (phytic acid) which inhibits the binding of L- and P-selectin to sialyl Lewis^X could inhibit the development of intimal hyperplasia. Endothelial injury was inflicted in one femoral artery via the photochemical reaction between systemically injected rose bengal and transillumination with green light (wavelength: 540 nm). Scanning electron microscopic observation at 3 days after the injury showed an increase in the number of leukocytes adhering to the injury site. Histological observation at 21 days showed that in the neutropenia group administered anti-neutrophil antibody and in the phytic acid-treated group the progression of intimal hyperplasia was significantly attenuated by comparison with the corresponding control groups.

These results suggest that neutrophil accumulation contributes to the initiation and/or development of intimal hyperplasia and L- and/or P-selectin may participate in their mechanisms.     

伊马替尼对兔颈动脉内膜增生的作用及机制研究

目的探讨伊马替尼对动脉球囊损伤后新生内膜的作用及机制。方法长耳白兔30只,按随机数字表法分为A、B、C组,分别于右侧颈动脉球囊拉伤后给予0、25、50 mg/kg伊马替尼灌胃1次/d,连续用药14 d处死。取双侧颈动脉行HE及免疫组化染色A组左侧颈动脉为对照组;逆转录PCR检测血小板衍生生长因子(PDGF)-B及受体PDGFR-βm RNA水平。ELISA法检测PDGF-BB血浆水平。结果 (1)术后新生内膜厚度、面积、内/中膜厚度及面积比值较对照组均增加(P<0.05)。(2)术侧组织PDGF-B m RNA水平A组高于B组及C组(2.961±0.686、1.957±0.545、1.879±0.636,F=8.353,P<0.01);PDGFR-βm RNA水平仅C组高于A组(1.236±0.356 vs 0.708±0.372,P<0.01)。(3)各组术后血浆PDGF-BB水平均高于术前(P<0.01),且A组>B组>C组(ng/L:23.464±3.542、19.504±2.454、16.588±1.207,F=17.322,P<0.05)。(4)血浆PDGF-BB水平与PDGF-B m RNA水平正相关(r=0.806,P<0.01)。结论伊马替尼通过抑制PDGF-B m RNA表达而抑制动脉内膜损伤后新生内膜过度增生。     

Sulfatides, L- and P-selectin ligands, exacerbate the intimal hyperplasia occurring after endothelial injury

Leukocytes may be important in the development of intimal hyperplasia, but little is known about the participation of sulfatides (3-sulfated galactosyl ceramides) which are native ligands of L- and P-selectin. This study was designed to determine whether sulfatides affect the development of intimal hyperplasia. ICR mice were randomized to receive vehicle or sulfatides intravenously either at 1, 3, or 10 mg/kg/day for 7 days, or at 10 mg/kg/day for 1, 3, or 7 days. Endothelial damage was inflicted on the femoral artery via the photochemical reaction between rose bengal and green light. Scanning electron and light microscopic observations 3 days after the injury indicated that sulfatides-treated animals had more neutrophils adhering to the injury site than vehicle-treated controls. At 21 days, sulfatides-treated animals had a greater neointimal area than controls. In in vitro studies, sulfatides (i) increased cytosolic free calcium in mouse neutrophils, (ii) caused increases in expression of Mac-1 (CD11b/CD18) on the neutrophil membrane surface in mouse whole blood. These findings suggest that neutrophil accumulation on the subendothelial matrix or adherence of platelets mediated by adhesive interactions between L- or P-selectin and sulfatides may contribute to the development of intimal hyperplasia. The neutrophil accumulation may be mediated by an increase in Mac-1 caused by the agonistic effects of sulfatides on the neutrophil membrane surface, or by an increase in L- and P-selectin ligands resulting from the binding of sulfatides onto the exposed subendothelial matrix.     

Effects of local cytochalasin D delivery on smooth muscle cell migration and on collar-induced intimal hyperplasia in the rabbit carotid artery

1. Smooth muscle cell (SMC) migration has been implicated in neointima formation after angioplasty. Therefore, we investigated whether cytochalasin D, a fungal metabolite that inhibits actin filament formation, suppressed SMC migration and collar-induced intimal hyperplasia in the rabbit carotid artery. 2. To establish effective concentrations, contractions of carotid artery rings to phenylephrine were determined after incubation with cytochalasin D (10(-8) - 10(-6) M) for 30 min or 3 days. In vitro cell migration was studied using carotid artery explants and a modified Boyden chamber with SMCs isolated from the rabbit aorta. The in vivo effect was tested after infusion of 10(-8) - 10(-4) M cytochalasin D into collars placed around the left carotid artery; collars placed around the right artery served as controls. 3. Contractions to phenylephrine decreased after 30 min or 3 days exposure to 10(-7) and 10(-6) M cytochalasin D; the effect was partly reversible. These concentrations also inhibited cellular outgrowth and SMC migration in the in vitro assays. 4. Immunohistochemistry showed that local delivery of 10(-5) or 10(-4) M cytochalasin D for 2 weeks suppressed collar-induced alpha-SMC actin expression in the intima by 68% and 84% respectively. However, the cross-sectional area of the intima was not reduced due to an influx of T-lymphocytes and macrophages. 5. It is concluded that cytochalasin D suppressed SMC contractility and migration in vitro. Although perivascular infusion of cytochalasin D inhibited collar-induced SMC migration from media to intima in vivo as well, the intimal hyperplasia was not reduced due to concomitant development of an inflammatory response.