目的:观察气体信号分子硫化氢(Hydrogen sulfide,H2 S)对乙醇所诱导的小鼠肾间质纤维化及结缔组织生长因子(CTGF)与基质金属蛋白酶(MMP)-8/基质金属蛋白酶组织抑制因子(TIMP)-1蛋白表达水平的影响.方法:将40只雄性小鼠随机分为对照组(Control组)、酒精性肾损伤组(ARD组)、硫化氢干预组(H2 S组)及乙醇加炔丙基甘氨酸组(PAG组).除Control组外,其他三组小鼠每天饮用4％(Vol/Vol)的乙醇水溶液12周以构建酒精性肾损伤模型;Control组每天饮用清水,H2S组每天腹腔注射硫氢化钠(H2S供体)溶液(50μmol/kg),PAG组每天腹腔注射硫化氢抑制剂PAG(40 mg/kg).12周后,处死小鼠取肾脏标本,PAS染色观察小鼠肾组织形态学变化和Masson染色观察小鼠肾脏组织间质纤维化改变,Western blot法检测CTGF、MMP-8和TIMP-1蛋白表达的变化.结果:与Control组相比,ARD组可见明显肾小管间质纤维化,肾小球和肾小管肿胀,同时CTGF、MMP8和TIMP1蛋白表达水平显著增高(P<0.05);与ARD组相比,H2S组肾损害程度减轻,肾小管间质纤维化明显改善,MMP-8、TIMP-1和CTGF蛋白表达水平也明显下调,差异有统计学意义(P<0.05),而PAG组差异无统计学意义(P>0.05).结论:硫化氢可改善乙醇所诱导小鼠肾小管间质纤维化,其机制可能与下调CTGF信号通路及其调控的MMP-8/TIMP-1有关.

目的 观察硫化氢(H2S)对阿霉素诱导的大鼠心肌纤维化及神经调节蛋白(NRG)1和ErbB2(又名HER2,NEU,CD340)蛋白表达的影响.方法 取SD雄性大鼠随机分为4组:对照组,模型组,H2 S模型组,H2 S对照组.经腹腔注射阿霉素建立心肌损伤大鼠模型,对照组和H2 S对照组经腹腔注射同等剂量的生理盐水;建模后,H2 S模型组和H2 S对照组每日经腹腔注射硫氢化钠溶液;Masson染色观察心肌纤维化及计算心肌胶原容积分数(CVF);Western印迹检测NRG1和ErbB2蛋白的表达.结果 与对照组相比,模型组心肌细胞紊乱,心肌纤维沉积明显增多、CVF显著增高(P<0.05),心肌NRG1和ErbB2蛋白的表达明显下降(P<0.05);而与模型组相比,H2S模型组心肌组织纤维沉积明显减少、CVF显著下降(P<0.05),NRG1和ErbB2蛋白的表达显著上调(P<0.05).结论 H2 S可以改善阿霉素导致的心肌纤维化,其发生机制可能与通过NRG1/ErbB2通路上调NRG1和ErbB2蛋白表达有关.

目的 探索通心络胶囊对压力超负荷大鼠心肌肥厚和QT离散度以及缝隙连接蛋白43(connexin 43,Cx43)表达的影响.方法 随机均分45只雄性SD大鼠成3组:对照组(control,C),模型组(model,M),治疗组(treat-ment,T),腹主动脉缩窄手术建立压力超负荷大鼠模型,予通心络胶囊[1.5 g/(kg·d)]干预12周后,以多普勒超声心动图检测,同时测定大鼠QT离散度,大鼠处死后取左室心肌组织行HE染色,以免疫组化方法检测Cx43的表达及分布,RT-PCR法检测Cx43 mRNA的表达.结果 M组大鼠心脏多普勒结果与C组比较显示IVSs、VPWs、LIVSd和LVPWd均有增加(均P＜0.01);而T组和M组比较,以上检测参数则得以改善.与C组相比,M组QT离散度明显延长(P＜0.01),与M组相比,T组QT离散度明显缩短(P＜0.01).同时HE染色显示:与C组相比,M组心肌细胞肥大明显(P＜0.01),呈无序排列,T组与M组比较,心肌肥大及排列均较其改善(均P＜0.01).RT-PCR及免疫组化检测结果均显示:与C组相比,M组Cx43 mRNA和蛋白的表达均有明显下调(均P＜0.05),而T组与M组相比,Cx43 mRNA和蛋白的表达有所上调(均P＜0.05).结论 通心络胶囊在改善压力超负荷大鼠心肌肥厚的同时也缩短QT离散度及上调Cx43的表达.

Objective:To investigate the influence of gefitinib on vascular smooth muscle cells (VSMCs) and endothelial cells (ECs) proliferation.Method:Cultured VSMCs and ECs were respectively treated with different concentrations and durations of gefitinib and paclitaxel.MTT method was used to measure the influence of these two drugs on VSMCs and ECs proliferation,then calculate the growth inhibition rate,respectively.Western blot was used to determine the expression of EGFR and p EGFR.Result:①Gefitinib and paclitaxel inhibited the proliferation of VSMCs and ECs in time-does ependence.Compared to paclitaxel,the same low concentrations of gefitinib had a comparable inhibition rate on smooth muscle cells at the same duration,but with the increase of concentration and extension of time,the inhibition rate of gefitinib increased significantly,and even exceeded paclitaxel.In addition,the inhibition of gefitinib on ECs was lower than paclitaxel with the same concentration in the same time period.②EGFR expressed in both of the two cell lines,but it expressed much higher in VSMC than in EC.Expression of p-EGFR in VSMCs was significantly suppressed after the role of gefitinib.Conclusion:Gefitinib and paclitaxel had a comparable suppression effects on VSMCs proliferation,the inhibition of Gefitinib group on ECs proliferation was obviously weaker than paclitaxel group.So it provided a theoretical and experimental basis for the application of Gefitinib in the treatment of restenosis and thrombosis after PCI.

目的:探讨表皮生长因子受体(epidermal growth factor receptor,EGFR)抑制剂吉非替尼对平滑肌细胞(smooth muscle cells,VSMC)和内皮细胞(ndothelial cells,EC)增殖的影响,以及对EGFR和Akt蛋白表达及磷酸化的影响.方法:将大鼠VSMC及EC置于含0.01～10μmol·L的吉非替尼的培养基中培养24～72 h,以MTT法测定细胞增殖的抑制率.Western blot检测EGFR及磷酸化EGFR(p-EGFR)、Akt及磷酸化Akt(p-Akt)蛋白水平.结果:MTT结果显示,吉非替尼抑制VSMC增殖呈时间和浓度依赖性,而吉非替尼对EC增殖的抑制作用明显低于紫杉醇;Western blot结果显示VSMC中EGFR(1.07±0.13)表达与EC(0.58±0.05)相比明显增多(P＜0.01),而吉非替尼可明显抑制VSMC中EGFR及Akt蛋白的磷酸化.结论:类似紫于杉醇,吉非替尼可抑制VSMC增殖,而对EC的细胞毒性作用明显低于紫杉醇,其机制可能是通过抑制EGFR及Akt蛋白磷酸化来实现的.

目的 通过心内科临床实习教学及考核中应用“模拟诊疗”,培养学生临床诊疗思维及独立工作能力.方法 100名学生随机分成两组,对照组采取常规心内科临床实习教学和考核；观察组采取“模拟诊疗”实习教学和考核,分析学生满意及兴趣度,比较两组学生临床诊疗能力,评价“模拟诊疗”考核不同环节的优缺点.结果 观察组学生实习满意度及兴趣度、医患沟通能力、病例分析技能、临床诊疗能力成绩明显优于对照组,差异有统计学意义(P＜0.05),“模拟诊疗”3环节各有优势特点,其中第2环节客观全面.结论 模拟诊疗考核激发学生学习兴趣,使技能考核的结果更加客观公正,使学生更明确实习的目的及学习的积极性；培养学生独立临床工作能力.

Aim To investigate the effects of hypoxia on the expression of Cx43 and apoptosis in hypertrophic rat cardiomyocytes.Methods Cardiomyocytes from newly born rats were cultured after digested in trypsin.The apoptosis rate of cardiomyocytes induced by hypoxia was evaluated by a modified Hoechst33258 fluorescent staining method.The expression of the Cx43 protein were detected by Western blot and fluorescent staining method.Results Compared with the control group,the apoptosis rate of hypertrophic cardiomyocytes induced by 24 h hypoxia was increased(15.42%±1.62%vs 2.56%±0.48%).The expression of Cx43 protein of hypertrophic cardiomyocytes was obviously decreased in comparison with the control group.Hypoxia could significantly down-regulate the Cx43 expression in hypertrophic cardiomyocytes than that of the normal cardiomyocytes(P<0.01).Conclusions Hypoxia can significantly increase the apoptosis rate and down-regulate the Cx43 expression in hypertrophic cardiomyocytes.These may be involved in the mechanisms of electrophysiology remodeling and malignant arrhythmia of hypertrophia myocardium induced by hypoxia.

Aim To investigate the effect of Shensongyangxin capsule on QT dispersion and connexin43 expression in heart failure rats. Methods Heart failure rats models were built by constricting abdominal aorta,and were lavaged with Shensongyangxin capsule for 8 weeks.Ventricular electrophysiology were measured by inserting home-made electrode into subcutaneous,left ventricular morphostructure,myocardial fibrosis,and connexin43 distribution were respectively observed by HE staining,Masson staining,immunohistochemical staining. Results Heart failure rats QT dispresion were significantly longer(37.20±9.94 ms,P<0.05),cardiomyocytes were misaligned,myocardial fibrosis area were significantly increased(101217.30±33970.02 μm2,P<0.05),and connexin43 distribution were significantly decreased(55.93±11.61,P<0.05).Shensongyangxin capsule can shorten QT dispresion(25.50±8.21 ms) of heart failure rats,increase connexin43 distribution(69.09±16.59) and decrease myocardial fibrosis area(13580.64±8213.73 μm2) in myocardium of heart failure rats. Conclusion Shensongyangxin capsule can shorten QT dispresion of heart failure rats,increase connexin43 distribution and decrease myocardial fibrosis area in myocardium of heart failure rats.

Objective To investigate the electrical instability and connexin 43(Cx43) expression changes in left ventricles of heart failure(HF) rats induced by pressure overload.Methods Surgical abdominal aortic stenosis was used to prepare pressure overload induced heart failure model in SD rats.The rats with left ventricular end-diastolic pressure(LVEDP)≥5 mmHg were categorized into HF group.Sham-operated rats served as controls.Alter 32 weeks,cardiac function of the rats was analyzed with carotid artery canalization and ultrosound B examination.The electrophysiological markers were also investigated.The changes in cardiac electrical instability and gap junction remodeling in left ventricles of rats were characterized by imniunoblotting and transmission electron microscopy.Results HF rats presented obvious cardiac electrical instability and cardiac dysfunction.The heart rate,QT-interval,LVEDP and ventricular effective refractory period(VERP) increased significantly,and left ventricular ejection fraction decreased significantly in HF group.The expression of Cx43 in left ventricle was decreased significantly(0.929±0.095 vs 1.250±0.083,P0.05) in HF group.Transmission electron microscopy revealed gap junction remodelling in HF rats.Conclusion Pressure overload induced HF rats present obvious gap junction remodelling in ventricles,which may be related to electrical instability in yentricles.

Objective To analysis the application value of DES and tirofiban to AMI patients.Methods 78 AMI patients were divided into observation group that were treated with DES and tirofiban and control group that were treated with DES.TIMI,MBG,drop of ST-segment,cardiac index and MACE were compared and analysised.Results Observation group's MBG,one week's drop of ST-segment after operation,cardiac index were better than control group's Obviously,observation group's rate of MACE was lower than control group's(7.5% Vs 23.7%).There were significant difference between the two groups(P <0.05).Conclusion Application of DES and tirofiban to AMI patients can make myocardial ischemia reperfusion,reduce no-reflow phenomena,reduce the incidence of heart attacks,can significantly improve patients' cardiac and prognosis.

Gap junctions are clusters of transmembrane channels that mediate direct exchange of ions and small signaling molecules between adjacent cells,connexin43 (Cx43)is the major gap junction protein expressed in cardiac muscle cells.Expression and spatial distribution of Cx43 is closely associated with arrhythmia,studies suggested that Cx43 might be a new target for treating arrhythmia.The relationship between Cx43 and arrhythmia and advances in the study of new therapy strategies based on Cx43 were reviewed in this article.

【Objective】 To investigate the inhibitory effects of human plasminogen kringle 5 (K5) on the growth and metastasis of mouse Lewis lung carcinoma (LLC) in vivo. 【Methods】 To evaluate K5 activity in vivo, the mouse lung cancer graft model was established by injection of Lewis lung carcinoma cells subcutaneously into the oxter of C57BL / 6J mice. On the 7th days after inoculation, the mice then received an intraperitoneal injection of K5 or phosphate buffered solution (PBS) as control. Tumor volume and mouse body weight were measured and plotted in tumor-volume curve and body-weight curve to estimate the tumor growth rate and general constitution of the animals during the course of treatment. A spontaneous metastasis mouse model of Lewis lung carcinoma was established by removing the primary tumors in the 7th days after inoculation. Right after the tumor remove, mice received K5 or PBS by intraperitoneal injection. The body weight and survival rate of both groups were recorded. After the mice were executed, the lungs were dissected, weighted, and undertaken a pathological analysis. 【Results】 The K5-treated group had a tumor growth rate which was significantly lower than that of the control group in grafted LLC mouse model, and distinctly depression of tumor weight [(4.57 ± 0.79) g, and (1.15 ± 0.31) g, respectively, P=0.006] and body weight. Compared to PBS-treated group, the incidence of metastasis to lung and the mean number of metastatic lung nodules (15.75 ± 9.79 and 6.60 ± 3.39 respectively, P=0.29) and the wet lung weight of each animal were reduced by K5 treatment in metastasis model of mouse Lewis lung carcinoma.Similarly, the pulmonary micrometastasis of K5-treated group decreased obviously with microscopic observation at high magnification. 【Conclusion】 K5 can inhibit the growth of Lewis lung carcinoma, and suppress the pulmonary metastasis of LLC cells in vivo.

Objective To study the effects of angiotensinⅡon expression of connexin43(Cx43) in cultured neonatal rat ventricular myocytes and cardiomyocyte hypertrophy.Methods Cardiomyocytes were isolated from newborn Wistar rats.AngiotensinⅡwas added into the media to induce myocyte hypertrophy.Cultures were exposed to 1 mmol/L angiotensinⅡfor 72 h.Cx43 expression was characterized by immunoblotting,RT-PCR and immunofluorescence methods.Hypertrophied myocytes was assessed by composition of protein assay and cell cycle phase assay.Results AngiotensinⅡin the range of 1.0×10~(-9)—1.0×10~(-6) mol/L induced hypertrophy of cardiomyocytes, increased the ratio of S and G_2—M phases cells,and concentration-dependently decreased Cx43-expression and its immunoreactivity by 37.6%(n=4,P0.05) as well Cx43 mRNA level(P0.05).Down-regulation of Cx43 were related with the changes of the ratio of cell cycle phase and related to its entrance into S phase of the cell cycle(r=0.912,P0.05).Conclusion AngiotensinⅡconcentration-dependently decrease in Cx43 mRNA and protein in cultured neonatal rat ventricular myocytes in association with hypertrophy of myocytes and cell growth cycle.

Objective To study the effects of angiotensin Ⅱ(AngⅡ),as a mediator of cardiac hypertrophy,on expression of connexin43(Cx43)in cultured neonatal rat ventricular myocytes.Methods Cardiomyocytes were isolated from newborn Wistar rats and divided randomly into the normal control group,the group treated with Ang Ⅱ(1.0×10-6mol/L)and the group treated with AngⅡ(1.0×10-6mol/L)plus valsartan(1.0×10-6mol/L).After the rat ventricular myocytes were treated with AngⅡ for different periods of time(12,24,48 and 72 h)and at different concentrations(1.0×10-9-1.0×10-5 mol/L),the changes of total amount of Cx43 protein in cultures exposed to AngⅡwere determined.Total protein was separated from control and treated cultures by SDS-PAGE and analyzed by immunoblotting.Meanwhile,the effect of valsartan on expression of Cx43 protein in cultures exposed to AngⅡ was observed by immunofluorescence technique.Results Immunofluorescence and immunoblotting analyses revealed that there were up-regulation of Cx43 protein in cultured neonatal rat ventricular myocytes treated with AngⅡ for 12 or 24 h and down-regulation of Cx43 protein in cultured cardiomyocytes treated with AngⅡ for 48 or 72 h.These changes were blocked by valsartan.The cultured neonatal rat cardiomyocytes exposed to increasing concentrations of AngⅡ(1.0×10-9-1.0×10-6 mol/L)for 72 h showed significant concentration-dependent decrease in Cx43 expression.Conclusion AngⅡ up-regulates expression of Cx43 protein in cultured neonatal rat ventricular myocytes treated for 12 or 24 h,and down-regulates Cx43 content after treated for 48-72 hours.These changes were concentration-dependent and can be blocked by valsartan,which can initiate remodeling of gap junctions.

AIM: To investigate the role of Rho-kinase signal pathway in rat cardiac fibroblasts (CFBs) proliferation and collagen synthesis induced by angiotensinⅡ (AngⅡ). METHODS: CFBs of neonatal Sprague-Dawley (SD) rats were isolated with the method of trypsin digestion and differential anchoring velocity. The CFBs were stimulated with AngⅡto induce fibrosis. Proliferation of CFBs was observed by MTT coloricmetric assay. Synthesis of collagen was detected by the hydroxyproline. The expression of Rho-kinase mRNA was examined using RT-PCR analysis. The extent of phosphorylation of myosin-binding subunit (MBS-P) of myosin phosphatase was quantified by Western blotting analysis, which was used to evaluate the activity of Rho-kinase.RESULTS: (1) Stimulation of neonatal SD rat CFBs with AngⅡ (10-7 mol/L) significantly increased CFBs proliferation and collagen synthesis (P0.01). (2)Stimulation of neonatal SD rat CFBs with AngⅡ (10-7 mol/L) significantly increased the expression of Rho-kinase mRNA and rapidly activated Rho-kinase in a time-dependent manner. (3) Within a concentration coverage, hydroxyfasudil (H4413), a Rho-kinase inhibitor, effectively inhibited AngⅡ-induced CFBs proliferation and collagen synthesis (P0.05 or P0.01).CONCLUSION: Rho-kinase signal pathway may be one of the most important signal transducter for AngⅡ-induced CFBs proliferation and collagen synthesis in neonatal SD rats.

Objective To investigate the characteristic and the mechanism of intimal thickening at different times after rat carotid arterial injury. Methods Thirty Anesthetized rats underwent right carotid arterial endothelial injury by balloon catheter introduced via the external carotid arterial. Rats were divided into five subgroups, which were killed at 1, 3, 9, 28, 60 days after balloon denudation, respectively.Carotid arteries were observed under micros and electron microscope and were analyzed by computerized morphometry. Results Irregular thickening of arterial intima and stenosis of arterial cavity were observed since 3 days after balloon denudation. At the 28th day the intimal thickening reached a peak. The proliferation characteristic at different times was related to the phenotype transformation of smooth muscle cells.Conclusion It indicates that intimal thickening after rat carotid arterial injury have a distinct time-dependent characteristic which is related to the phenotype transformation of smooth muscle cells.

目的探讨生物导向药物TGFα-SAP对平滑肌细胞增殖及动脉损伤后内膜增生的特异性抑制作用.方法用SPDP化学联结的方法合成TGFα-SAP,采用细胞计数方法观察TGFα-SAP对培养中的增殖平滑肌细胞的细胞毒性作用,并以3H-leucine渗入法进一步了解TGFα-SAP对平滑肌细胞的蛋白质合成的影响;在体实验中将Sprague-Dawle大鼠随机分为治疗组和对照组,均行右颈总动脉球囊内膜剥脱术,治疗组术后局部注射TGFα-SAP,对照组予以生理盐水;于不同时间点处死动物取动脉段行光镜观察及计算机图像分析.结果从体外实验可见TGFα-SAP能明显抑制SMCs的生长增殖及蛋白质合成;在体实验中图像分析结果显示:动脉损伤后TCFα-SAP治疗组在第3,9和28天内膜/内膜加中膜面积显著小于对照组(P＜0.05).结论TGFα-SAP与Saporin相比对增殖平滑肌细胞具有明显增强的细胞毒性,在体实验中亦可见其对损伤后的内膜增生具有明显的抑制作用.