TGF-β1/Smad2通路参与IL-17诱导的A549细胞上皮-间质转化

目的:探讨白细胞介素(IL)-17对A549细胞上皮-间质转化的影响及TGF-β1/Smad2信号通路在其中的作用。方法:用不同浓度(0,10,25,50,100ng/ml)IL-17刺激A549细胞48h,RT-PCR检测E-钙黏蛋白(E-cad)、α-平滑肌肌动蛋白(SMA)、TGF-β1的表达。将体外培养的细胞分为3组:对照组、IL-17(100ng/ml)组、IL-17+SB431542组。48h后收集各组细胞,RT-PCR和Western Blot检测E-cad、α-SMA、TGF-β1、Smad2、和p-Smad2的表达。结果:不同浓度的IL-17作用后,E-cad在50ng/ml IL-17组时表达下调,与浓度呈负相关;α-SMA在100ng/ml IL-17组时表达上调;TGF-β1在25ng/ml IL-17组时表达上调,与浓度呈正相关。与对照组相比,IL-17组E-cad表达下调,α-SMA、TGF-β1、p-Smad2表达均增加,Smad2蛋白表达水平无明显变化;与IL-17组比较,IL-17+SB431542组Smad2蛋白表达水平无明显变化,TGF-β1、p-Smad2、α-SMA蛋白表达均降低,E-cad的表达明显增加。结论:TGF-β1/Smad2通路参与IL-17诱导的A549细胞上皮-间质转化。     

TGF-β1诱导Ⅱ型肺泡上皮-间充质转化及ERK1/2信号系统的变化

目的探讨转化生长因子β1（TGF-β1）对Ⅱ型肺泡上皮-间充质转化的作用及与胞外调节激酶1/2（ERK1/2）信号系统的关系。方法体外培养人Ⅱ型肺泡上皮细胞来源的A549细胞,分为阴性对照组和TGF-β1处理组（0.05、0.5、5、10μg/L,作用时间48 h）。免疫荧光及Western blot方法检测各组细胞E钙粘素（E-cad）、α-平滑肌肌动蛋白（α-SMA）、波形蛋白（Vim）和纤维连接蛋白（Fn）的表达。在TGF-β1作用不同时间,采用Western blot及RT-PCR方法检测各组细胞α-SMA、E-cad、Vim、Fn、ERK1/2和p-ERK1/2的表达。结果与阴性对照组比较,TGF-β1组（5μg/L）α-SMA、Vim、Fn蛋白表达显著升高,而E-cad表达显著降低（P〈0.05）。作用6 h后,TGF-β1组出现α-SMA mRNA表达显著升高,E-cad显著下降（P〈0.05）。作用48 h后,TGF-β1组出现Vim、Fn蛋白表达显著升高,E-cad显著下降（P〈0.01）。作用30 min后,TGF-β1组ERK/2磷酸化水平显著升高（P〈0.05）。结论 TGF-β1可在体外以浓度和时间依赖方式的诱导A549细胞发生上皮-间充质转化,其机制可能与上调ERK1/2信号转导通路有关。     

TGF-β信号通路及细胞黏附分子在非小细胞肺癌的研究

目的观察TGF-β1、TβRⅡ、Smad2、E-cad、E-sel、integrin-β1、ICAM-1及CD44v6在非小细胞肺癌（NSCLC）中的表达特征,探讨其与非小细胞肺癌的组织学类型、TNM分期及淋巴结转移的关系。方法采用免疫组织化学SP和SABC法检测TGF-β1、TβRⅡ、Smad2、E-cad、E-sel、integrin-β1、ICAM-1及CD44v6的表达,并对其与NSCLC临床病理因素的关系进行分析。结果TGF-β1与integrin-β1在肺腺癌组织的阳性表达率显著高于肺鳞状细胞癌组织（P〈0.05）,E-cad在肺腺癌组织的阳性表达率显著低于肺鳞状细胞癌组织（P〈0.05）。TGF-β1及TβRⅡ在NSCLC TNM分期Ⅰ期组织的阳性表达率显著低于Ⅱ期和Ⅲ期组织（P〈0.01）。TGF-β1、TβRⅡ、Smad2及CD44v6在有淋巴结转移的NSCLC组织中阳性表达率显著高于无淋巴结转移的NSCLC组织（P〈0.05）,E-cad在有淋巴结转移的NSCLC组织中阳性表达率显著低于无淋巴结转移的NSCLC组织（P〈0.05）。结论TGF-β1、E-cad及integrin-β1与NSCLC的组织学类型有关,TGF-β1及TβRⅡ能促进NSCLC的病程进展;TGF-β1、TβRⅡ、Smad2、E-cad、及CD44v6的阳性表达与NSCLC的淋巴结转移密切相关,TGF-β1、TβRⅡ、Smad2、E-cad及CD44v6在NSCLC的淋巴结转移中起着重要的促进作用,可作为估计NSCLC可能有淋巴结转移的指标。     

金氏补肾活血方对卵巢储备功能减退大鼠TGF-β1/Smads信号通路及性激素的影响

目的 观察金氏补肾活血方对卵巢储备功能减退模型大鼠的性激素、卵巢组织形态的作用及其调节转化生长因子-β1(TGF-β1)/Smads信号通路的可能机制。方法 将50只SD雌性大鼠随机分为5组:空白组、模型组、西药组、中药低剂量组、中药高剂量组,每组10只。除空白组外,其余各组均给予环磷酰胺75 mg/kg腹腔注射,复制卵巢储备功能减退(DOR)大鼠模型。经灌胃治疗后,阴道涂片观察大鼠动情周期变化、酶联免疫法(ELISA)检测血清性激素、苏木素-伊红(HE)染色观察卵巢组织形态、荧光定量聚合酶链反应法(RT-qPCR)检测大鼠卵巢转化生长因子-β1(TGF-β1)、Smad2、Smad3、Smad7的基因表达。结果 与空白组比较,膜型组大鼠的动情周期丧失规律性,血清雌二醇(E₂)、抗缪勒管激素(AMH)水平显著降低(P<0.01),促卵泡激素(FSH)、促黄体生成素(LH)水平显著升高(P<0.01),卵巢指数、子宫指数明显降低(P<0.05),卵泡发育出现闭锁,卵巢中TGF-β1、Smad2、Smad3的表达量均减少(P<0.01),Sma...     

基于TGF - β/smad信号通路探讨七方胃痛颗粒对人胃腺癌AGS细胞上皮间质转化的影响

目的:基于TGF-β/smad信号通路探讨七方胃痛颗粒对人胃腺癌AGS-EMT细胞ZEB2、E-cad及Vimentin的影响及可能的机制。方法:用TGF-β1处理AGS细胞建立上皮间质转化(epithelial mesenchymal transition,EMT)模型,将细胞分为空白组、七方胃痛颗粒(含药血清+EMT模型)组、LY2109761(TGF-β/Smad抑制剂+EMT模型)组,用RT-PCR、Western Blot法检测ZEB2、E-cad及Vimentin的表达。结果:与空白组比较,七方胃痛颗粒及LY2109761干预后能上调E-cad蛋白及其mRNA的表达,下调ZEB2蛋白及其mRNA和Vimentin蛋白及其mRNA的表达(P<0.05),并呈时间依赖性。与七方胃痛颗粒组相比,LY2109761组ZEB2蛋白及其mRNA的表达明显下调(P<0.05)。结论:七方胃痛颗粒与LY2109761作用相似,其可能是通过抑制TGF-β1/Smad信号通路,上调E-cad表达,下调ZEB2及Vimentin表达,抑制EMT形成,从而抑制人胃腺癌AGS细胞的上皮-间质转化。     

姜黄素通过TGF-β/Smads信号通路对肾缺血再灌注后肾小管间质纤维化抑制作用

目的:探究姜黄素通过TGF-β/Smads信号通路对肾缺血再灌注后肾小管间质纤维化的抑制作用.方法:将40只健康SD大鼠按随机数字表法平均分为假手术组、模型组、低剂量组和高剂量组.使用夹闭法松开肾蒂的方法复制肾缺血再灌注大鼠模型.检测血清中血清肌酐(serum creatinine,Scr)和尿素氮(urea nitrogen,BUN)水平、肾组织胶原含量、肾组织的病理变化以及肾脏中转化生长因子-β1 (transforming growth factor β1,TGF-β1)、Smad3和Smad7蛋白表达水平.结果:假手术组肾小管形态正常,模型组肾小管间质呈现明显的病理变化,低剂量组和高剂量组肾小管间质纤维病理变化明显改善,且高剂量组改善程度高于低剂量组;模型组的肾小管间质纤维化评分、Scr、BUN、胶原含量、TGF-β1和Smad3明显高于假手术组,Smad7明显低于假手术组(P＜0.05);低剂量组和高剂量组肾小管间质纤维化评分、Scr、BUN、胶原含量、TGF-β1和Smad3均明显低于模型组,Smad7明显高于模型组(P＜0.05);高剂量组肾小管间质纤维化评分、Scr、BUN、胶原含量、TGF-β1和Smad3评分明显低于低剂量组,Smad7明显高于低剂量组(P＜0.05).结论:姜黄素对肾缺血再灌注后肾小管间质纤维化具有抑制作用,其作用机制可能为抑制TGF-β/Smad信号转导通路.     

TGF-β1与胃肠道间质瘤预后关系研究

目的 探讨胃肠道间质瘤(GIST)中TGF-β1的表达与其顸后及机制的可能关系.方法 收集湘雅医院2001年12月～2005年12月77例经手术治疗GIST患者临床资料及手术病理标本,应用免疫组织化学(SP)法观察各组胃肠道间质瘤中TGF-β1、E-钙黏素(E-cadherin,E-cad)及N-钙黏素(N-cadherin,N-cad)的表达情况.结果 77例GIST中有38例TGF-β1表达阳性,47例E-cad表达阳性,37例N-cad表达阳性,阳性表达率分别为49.3%、61.0%和48.1%.TGF-β1的表达与组织学分级有关(P＜0.05).患者TGF-β1阳性组和阴性组5年生存率分别为49.0%和72.9%,两者生存率相比有显著性差异(P＜0.05),TGF-β1和E-cad的表达呈负相关性(r=-0.490,P＜0.05),TGF-β1与N-cad的表达呈正相关(r=0.610,P＜0.05).结论 TGF-β1随着GIST侵袭程度的增高其表达有所增加,TGF-β1可能是影响胃肠道间质瘤的预后的一个比较重要的因素.TGF-β1可能通过促进肿瘤发生类似上皮间质转化使肿瘤预后不佳.     

间质流膏治疗特发性间质性肺炎30例

目的观察间质流膏治疗特发性间质性肺炎(idiopathic interstitial pneumonia,IIP)的临床疗效。方法将60例IIP患者随机分为间质流膏组与激素组,分别采用间质流膏、强的松治疗12周,观察患者治疗前后临床、影像、生理评分(clinical,radiologic,and physiological score,CRP)及血清转化生长因子-β1(trans-forming growth factor-β1,TGF-β1)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的改变。结果两组治疗后CRP评分及TGF-β1、TNF-α水平均较治疗前显著下降(P<0.01)。间质流膏组CRP评分及TGF-β1、TNF-α水平均显著低于激素组(P<0.01)。结论间质流膏较单纯激素治疗可有效改善IIP患者临床症状,提高生存质量,并能显著降低患者血清TGF-β1、TNF-α水平。     

二黄益肾汤调控TGF-β1/Smad信号通路治疗慢性肾功能衰竭研究

[目的] 观察二黄益肾汤对5/6肾切除大鼠肾组织血清尿素氮（BUN）、血清肌酐（Scr）、转化生长因子-β1（TGF-β1）、Smad3、Smad4、Smad7 mRNA及蛋白表达的影响。[方法] 健康SPF级雄性SD大鼠,采用5/6肾切除方式建立慢性肾功能衰竭模型。造模成功后,随机分为模型组、尿毒清组和二黄益肾汤组,各给药组分别给予相应药物进行灌胃治疗,时间为12周。实验结束后,检测BUN、Scr含量,苏木精-伊红（HE）染色分析病理改变。采用实时定量-聚合酶链反应（RT-PCR）和蛋白免疫印迹（Western Blot）方法检测TGF-β1、Smad3、Smad4和Smad7 mRNA及蛋白表达量。[结果] 5/6肾切除方法可以成功建立慢性肾功能衰竭动物模型。与模型组比较,二黄益肾汤组与尿毒清组能够降低Scr、BUN含量,差异具有统计学意义（P<0.01）。二黄益肾汤组和尿毒清组肾间质内有少量炎性细胞浸润及纤维组织增生,纤维化面积缩小,能够改善肾功能衰竭肾纤维化病变程度。基因和蛋白表达量分析结果显示,与模型组比较,二黄益肾汤组和尿毒清组TGF-β1、Smad3、Smad4表达呈降低趋势,差异有统计学意义（P<0.05）,Smad7的表达呈升高趋势,差异有统计学意义（P<0.05）。[结论] 二黄益肾汤能够改善慢性肾功能衰竭大鼠肾脏纤维化程度,抑制TGF-β1、Smad3、Smad4的基因和蛋白表达量,促进Smad7的表达量增多,可能通过调控TGF-β1/Smad信号通路而发挥相关治疗作用。     

桔梗皂苷D基于TGF-β/Smad通路调控上皮-间质转化对人子宫内膜癌Ishiwaka细胞迁移及侵袭的作用

目的 探讨桔梗皂苷D调控上皮间质转化（EMT）对人子宫内膜癌Ishiwaka细胞迁移、侵袭的影响,及对转化生长因子β（TGF-β）/Smad信号通路的调节作用。方法 取第4代对数生长期的人子宫内膜癌Ishiwaka细胞,随机分为对照组、桔梗皂苷D组、激动剂组和桔梗皂苷D+激动剂组。桔梗皂苷D组细胞加入桔梗皂苷D 18 μmol/L培养24 h,激动剂组细胞加入重组人TGF-β 20 ng/mL刺激24 h,桔梗皂苷D组+激动剂组细胞先加入重组人TGF-β 20 ng/mL刺激24 h后,再加入桔梗皂苷D培养24 h。MTT法检测细胞存活率,Transwell实验检测细胞侵袭能力,划痕实验检测细胞迁移能力及蛋白印迹法检测TGF-β1、p-smad2/3、上皮钙粘蛋白（E-cad）和神经钙粘蛋白（N-cad）相对表达量。结果 与对照组比较,桔梗皂苷D组细胞存活率及TGF-β1、p-smad2/3和N-cad蛋白相对表达量降低,细胞迁移和侵袭能力减弱,E-cad蛋白相对表达量升高,激动剂组TGF-β1、p-smad2/3和N-cad蛋白相对表达量升高,细胞迁移和侵袭能力增强,E-cad蛋白相对表达量降低（P＜0.05）;与桔梗皂苷组比较,桔梗皂苷D+激动剂组细胞存活率及TGF-β1、p-smad2/3和N-cad蛋白相对表达量升高,细胞迁移和侵袭能力增强,E-cad蛋白相对表达量降低（P＜0.05）;与激动剂组比较,桔梗皂苷D+激动剂组细胞存活率及TGF-β1、p-smad2/3和N-cad蛋白相对表达量降低,细胞迁移和侵袭能力减弱,E-cad蛋白相对表达量升高（P＜0.05）。结论 桔梗皂苷D可能通过抑制TGF-β1/Smad2/3信号通路调控上皮间质转化降低人子宫内膜癌Ishiwaka细胞迁移及侵袭能力。     

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.     

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.     

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…     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

CLINICAL SIGNIFICANCE OF SERUM CYTOKINES IL-1β,sIL-2R,IL-6,TNF-α,AND IFN-v IN ACUTE CORONARY SYNDROME

Objectives To explore serum cytokines levels (including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v) and their significance in patients with acute coronary syndrome (ACS) and the subsequent follow-ups, with attempt to estimate the role of various serum inflammatory markers in the diagnosis and assessment of ACS.Methods The study population include 40 patients with acute myocardial infarction (AMI), 40 patients with unstable angina pectoris (UAP), and 40 controls. Among the 80 patients, 60 patients attended a follow up 4 months later. Serum inflammatory markers including IL-1 β, sIL-2R, IL-6, TNF-α, and IFN-v were measured by enzyme linked immunosorbent assay.Results Serum IL- 1 β, sIL-2R, IL-6, TNF-α were significantly higher in AMI group or UAP group compared to the control group and became significantly lower 4 months later in the follow-up patients. Serum levels of IFN-v shows no significant difference between AMI group or UAP group and controls, also showing no significant change when measured in follow up patients. There was no correlation between serum creatine kinase-MB isoenzyme levels and serum inflammatory markers either in UAP or AMI group. Furthermore, when divided into two subgroups using Wagner's QRS scoring system in the AMI group, there is no difference of each serum inflammatory marker between ≤ 6 scores group and ＞ 6 scores group.Conclusion Serum levels of certain inflammatory markers may have some diagnostic value for ACS, and can be a useful marker reflecting disease stability.