线粒体ALDH2对高糖处理的乳鼠心肌成纤维细胞中自噬相关蛋白影响

目的探讨自噬是否介导乙醛脱氢酶（ALDH）2对高糖处理的乳鼠心肌成纤维细胞的作用。方法无菌条件下取出生3d内 SD大鼠乳鼠心脏,剪碎并用胰蛋白酶和胶原酶2混合酶消化成单个细胞,经差速贴壁后并传代培养,当传至3代时进行处理。应 用免疫荧光检测培养的乳鼠心肌成纤维细胞内波形蛋白以此鉴定原代培养心肌成纤维细胞纯度;实验涉及如下分组：5.5 mmol/L 糖对照组（F）、30 mmol/L高糖组（FH）、30 mmol/L高糖加乙醛脱氢酶2激动剂（Alda-1）组(FHA)、30 mmol/L高糖加乙醛脱氢酶2 抑制剂（Daidzin）组（FHD）、30 mmol/L高糖加乙醛脱氢酶2激动剂（Alda-1）和乙醛脱氢酶2抑制剂（Daidzin）（FHAD）;Western blot检测ALDH2、微管相关蛋白1轻链3B亚基（LC3B）、Beclin-1的蛋白表达;羟脯胺酸试剂盒测定细胞培养液中羟脯胺酸含量; CCK-8试剂盒测定心肌成纤维细胞增殖能力。结果与F组相比,FH组ALDH2蛋白表达降低,Beclin-1、LC3B蛋白表达降低,细 胞培养上清液内羟脯胺酸含量、细胞数目增高;与FH组相比,FHA组Beclin-1、LC3B蛋白表达、ALDH2蛋白表达增加,细胞数 目、细胞内羟脯胺酸含量降低,与此同时FHD组ALDH2蛋白及Beclin-1、LC3B表达降低,细胞数目、细胞内羟脯胺酸含量增高。 结论线粒体ALDH2抑制高糖诱导的乳鼠心肌成纤维细胞增殖作用,其机制可能与ALDH2对自噬相关蛋白Beclin-1、LC3B的 上调作用有关。     

肾上腺髓质素对醛固酮促心肌成纤维细胞增殖的影响

目的:探讨肾上腺髓质素(ADM)对醛固酮(ALDO)促心肌成纤维细胞增殖的影响及丝裂原激活的蛋白激酶(MAPK)信号转导途径的作用。方法:分离培养SD乳鼠心肌成纤维细胞,将细胞分成对照组、不同浓度ALDO组、ALDO+不同浓度ADM组、ALDO+ADM受体拮抗剂组,用3H-胸腺嘧啶([3H]-TdR)掺入和MAPK活性反映心肌成纤维细胞增殖。结果:ALDO呈浓度依赖性增加心肌成纤维细胞3H-TdR掺入,ALDO强烈刺激成纤维细胞ADM分泌和ADMmRNA表达,亦呈浓度依赖性,加入ADM(10-9～10-7mol/L)呈浓度依赖性抑制ALDO促心肌成纤维细胞增殖作用,其[3H]-TdR掺入量减低,抑制率分别为16.8%-37.4%(P<0.01),其MAPK活性明显减低,抑制率为7.4%-33.6%(P<0.05或P<0.01)。ADM22-52明显增强ALDO的促心肌成纤维细胞3H-TdR掺入效应,而CGRP8-37对ALDO的促细胞增殖效应无明显影响。结论:ADM对ALDO促心肌成纤维细胞增殖具有抑制作用,同时具有下调MAPK活性的作用,ADM对心肌成纤维细胞增殖的抑制效应可能由MAPK信号途径所介导。     

高糖加高胰岛素对心肌成纤维细胞增殖的影响

目的 利用体外培养的乳鼠心肌成纤维细胞，观察高浓度葡萄糖加高浓度胰岛素对心肌成纤维细胞增殖的影响，并探讨其可能作用机制。方法 以培养的乳鼠心肌成纤维细胞为模型分组给药后，利用结晶紫染色法观察细胞的增殖情况；利用[^3H]thymidine标记法测定细胞DNA的合成；利用流式细胞仪分析细胞周期。结果与正常对照组相比，高糖、高胰岛素、高糖加高胰岛素组分别造成了心肌成纤维细胞数目、DNA合成及S＋G2＋M期细胞的百分比增加，（P〈0．01），而高糖加高胰岛素组的各项指标值显著高于其它各组，（P〈0．01）。结论 高糖加高胰岛素具有协同促进心肌成纤维细胞增殖的作用。     

氧化还原因子1促进乳鼠心肌成纤维细胞增殖

目的：研究氧化还原因子1 (redox factor 1, REF1)对乳鼠心肌成纤维细胞增殖和胶原合成的影响及其可能的机制。方法：为证实REF1在乳鼠心肌成纤维细胞增殖和胶原合成中的作用,构建重组Ref1野生型和突变型的腺病毒载体,感染乳鼠心肌成纤维细胞。通过RT-PCR和Western 印迹检测Ref1以及与胶原合成相关的基因表达的改变,通过免疫荧光检测REF1核转位,利用MTT和流式细胞仪分析细胞增殖和细胞周期,用电泳迁移率实验(electrophoretic mobility shift assay,EMSA)检测REF1对转录因子活化蛋白1 (activator protein 1, AP1)的DNA结合能力的影响。利用含25 mmol/L的葡萄糖培养基模拟高糖环境,观察对乳鼠心肌成纤维细胞增殖及REF1表达与活性的影响。结果： MTT比色法显示,感染野生型Ref1重组腺病毒的细胞在490 nm处的光密度值明显高于对照病毒组（0.671±0.044 vs 0.364±0.007,n=6,P<0.01）。流式细胞仪对细胞周期的分析表明,野生型Ref1能明显增加S期细胞的百分率（16.8%±0.62% vs 9.04%±0.43%,n=3,P<0.05）。RT PCR检测发现,野生型Ref1促进细胞Ⅰ型胶原(collagen I, Col Ⅰ)和Ⅲ型胶原(collagen Ⅲ, Col Ⅲ) mRNA表达增多,但突变型Ref1对细胞的增殖和胶原的合成均没有明显影响。EMSA实验结果显示,感染野生型Ref1重组腺病毒的细胞AP1的DNA结合能力增强。高糖培养基培养乳鼠心肌成纤维细胞同样能增强AP1的DNA结合能力。高糖培养虽然对Ref1的表达水平没有影响,但增强了细胞中REF1的核转位,促进了细胞的增殖。结论：REF1促进乳鼠心肌成纤维细胞的增殖和胶原合成,其机制可能是通过上调AP1的DNA结合能力。     

TNF-α对培养乳鼠心肌成纤维细胞的作用

目的:探讨不同浓度TNF-α对培养乳鼠心肌成纤维细胞增殖、分泌AngⅡ、胶原及AT1受体表达的影响。方法:原代培养SD乳鼠心肌成纤维细胞,采用MTT法检测心肌成纤维细胞增殖,羟脯氨酸测定法观察成纤维细胞培养上清胶原含量,酶免法检测心肌成纤维细胞培养液AngⅡ含量,免疫细胞化学法观察心肌成纤维细胞膜AT1受体的表达变化。结果:TNF-α呈浓度依赖性的促进培养的乳鼠心肌成纤维细胞分泌AngⅡ和胶原,促进心肌成纤维细胞增殖及AT1受体的表达。结论:TNF-α促进心肌成纤维细胞分泌AngⅡ介导了心肌纤维化而参与心肌改建。     

人参皂苷Rg1对高糖培养的心肌成纤维细胞Wnt信号通路的影响

目的 建立体外高糖培养心肌成纤维细胞(CFs)的方法,观察人参皂苷Rg1对高浓度葡萄糖培养的心肌成纤维细胞增殖和蛋白分泌及Wnt信号通路的影响.方法 采用体外高糖诱导心肌成纤维细胞增殖模型,应用MTT法检测细胞增殖能力;流式细胞仪检测细胞增殖周期;ELISA法观察细胞Ⅰ型、Ⅲ型胶原及肿瘤生长因子β1(TGF-β1)蛋白的分泌;Western blot法检测β-catenin、糖原合成酶激酶3β(GSK-3β)、磷酸化糖原合成酶激酶3β(P-GSK-3β)的蛋白表达水平以观察人参皂苷Rg1对于心肌成纤维细胞Wnt信号通路的影响.结果 人参皂苷Rg1可以抑制细胞增殖(P＜0.05);降低Ⅰ型、Ⅲ型胶原分泌(P＜0.05);减少TGF-β1的合成(P＜0.01);在分子水平上可以抑制β-catenin的表达(P＜0.05),上调P-GSK-3β的表达(P＜0.05).结论 人参皂苷Rg1能抑制心肌成纤维细胞的增殖,减少高糖培养心肌成纤维细胞胶原和TGF-β1的分泌,抑制Wnt信号通路的异常表达,具有潜在的抗心肌纤维化作用.     

阿魏酸钠抑制心肌成纤维细胞增殖的作用及机制

目的:研究阿魏酸钠(SF)对AngⅡ诱导的心肌成纤维细胞(CFs)增殖的作用及其机制。方法:培养新生鼠心肌成纤维细胞分为3组:对照组、AngⅡ(10-7mol/L)组和AngⅡ+SF组。MTT法检测细胞增殖;流式细胞仪分析细胞周期改变;免疫荧光法检测TGFβ1蛋白表达。结果:AngⅡ作用24h后MTTOD值显著高于对照组(P<0.01)。不同浓度SF(0.04mg/ml,0.08mg/ml,0.16mg/ml)组MTTOD值均显著低于AngⅡ组(P<0.01),呈剂量依赖性。AngⅡ组S期细胞百分率明显高于对照组,G0/G1期细胞百分率则明显低于对照组(P<0.01)。AngⅡ+SF组S期细胞百分率显著低于AngⅡ组,G0/G1期细胞百分率显著高于AngⅡ组(P<0.01),且呈剂量依赖性。AngⅡ组TGFβ1蛋白表达显著高于对照组(P<0.01),AngⅡ+SF组显著低于AngⅡ组(P<0.01)。结论:阿魏酸钠可抑制AngⅡ诱导的心肌成纤维细胞增殖,其效应呈现剂量依赖性,作用机制与降低TGFβ1蛋白表达有关。     

ALK7在高糖诱导的心肌成纤维细胞转化及Ⅰ型胶原合成中的作用

目的 研究高糖诱导的心肌间质纤维化(心肌成纤维细胞表型转化以及胶原分泌)过程中活化素受体样激酶7(ALK7)的作用。方法 体外高糖培养大鼠心肌成纤维细胞,免疫荧光法观察成纤维细胞表型转化, real time PCR法检测心肌成纤维细胞ALK7和Ⅰ型前胶原mRNA的表达水平,ELISA法检测心肌成纤维细胞分泌Ⅰ型胶原水平,Western blotting法测定心肌成纤维细胞ALK7蛋白的表达水平;siRNA抑制ALK7表达,再次观察高糖对心肌成纤维细胞表型转化及上述各因子表达的影响。结果 高糖刺激心肌成纤维细胞转化为肌成纤维细胞;高糖促进Ⅰ型胶原mRNA 和蛋白的表达(P＜0.05);高糖促进成纤维细胞ALK7 mRNA和蛋白的表达(P＜0.05); ALK7 siRNA 抑制ALK7的表达后,高糖促进心肌成纤维细胞转化及合成Ⅰ型胶原的作用下降(P＜0.05)。结论 ALK7参与了高糖诱导的心肌成纤维细胞表型转化及合成Ⅰ型胶原的代谢过程。     

瓜蒌薤白半夏汤对心肌纤维化中整合素β1的抑制作用

摘 要：目的 研究整合素 β1 和纤维连接蛋白在血管紧张素Ⅱ（AngⅡ）诱导心肌纤维化中的作用,探讨瓜蒌薤白半夏汤(GXB)抗心肌纤维化的作用机制。方法 采用胰酶消化法培养乳鼠心肌成纤维细胞,建立 AngⅡ 诱导心肌纤维化模型。实验分为空白对照组,AngII 组,5%、10%、15% GXB含药血清组和缬沙坦含药血清组。空白对照组心肌成纤维细胞以空白对照组大鼠血清温育,其余各组均以 AngⅡ 1×10-6 mmol/L 刺激 24 h 后,再分别以空白对照组大鼠血清、5% GXB含药血清、10% GXB含药血清、15% GXB含药血清、缬沙坦含药血清温育 24 h。RT-PCR 法检测GXB对整合素 β1 mRNA 表达水平的影响;ELISA 检测培养上清液中纤维连接蛋白的量;MTT 法检测心肌成纤维细胞增殖。结果 与空白对照组比较,AngⅡ 1×10-6 mmol/L 作用心肌成纤维细胞 24 h,整合素 β1 mRNA 表达水平及纤维连接蛋白的量明显上调（P＜0.01）,心肌成纤维细胞增殖指数明显升高（P＜0.01）;经GXB含药血清干预后,与AngII组比较,整合素β1 mRNA的表达水平明显下降（P＜0.01）,纤维连接蛋白的量减少（P＜0.05）,心肌成纤维细胞增殖指数明显降低（P＜0.01）。结论 瓜蒌薤白半夏汤能够抑制 AngⅡ 诱导心肌纤维化,其作用机制可能与下调纤维连接蛋白/整合素β1表达有关。     

高糖状态下心肌成纤维细胞Ⅰ、Ⅲ型胶原表达变化的观察

目的探讨高糖对大鼠心肌成纤维细胞Ⅰ、Ⅲ型胶原表达的影响。方法采用胰酶消化法和差速贴壁分离法获取心肌成纤维细胞,采用正常糖(5.5mmol/L,对照组)和高糖(25mmol/L,高糖组)干预。用实时荧光定量PCR(Real-time PCR)方法检测Ⅰ、Ⅲ型前胶原mRNA表达,酶联免疫吸附测定法(ELISA)观察Ⅰ、Ⅲ型胶原蛋白水平的变化。结果与对照组比较,高糖刺激12h可使心肌成纤维细胞Ⅰ、Ⅲ型前胶原mRNA表达增加,Ⅰ、Ⅲ前胶原mRNA表达在12h升高,在24h达高峰,Ⅰ型前胶原mRNA升高幅度高于Ⅲ型前胶原mRNA;高糖刺激48h可使Ⅰ、Ⅲ型胶原蛋白表达增加,Ⅰ型胶原蛋白升高幅度高于Ⅲ型胶原。结论高糖可促进大鼠心肌成纤维细胞合成Ⅰ、Ⅲ型胶原。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.