G蛋白偶联受体激酶4对大鼠血管平滑肌细胞AT1受体的调节作用

目的应用siRNA干扰抑制大鼠胸主动脉血管平滑肌细胞G蛋白偶联受体激酶4(G protein-coupled recep-tor kinase 4,GRK4)的表达,探讨GRK4对血管紧张素Ⅱ1型(angiotensinⅡtype 1,AT1)受体的调节作用。方法免疫组化检测大鼠回结肠动脉血管平滑肌组织GRK4蛋白表达;以大鼠胸主动脉平滑肌细胞株(A10细胞株)为研究对象,免疫印迹检测GRK4、AT1受体蛋白表达变化,免疫共沉淀检测GRK4和AT1受体的相互作用和AT1受体磷酸化改变。结果大鼠动脉平滑肌组织GRK4表达良好;siRNA干扰后,GRK4蛋白表达明显下降(P<0.05);AT1蛋白表达降低(P<0.05),AT1受体磷酸化明显增强(P<0.05);GRK4和AT1受体存在共连接,抑制GRK4表达后增加GRK4与AT1受体之间的共连接。结论 GRK4能够调控大鼠胸主动脉平滑肌细胞AT1受体蛋白表达及其磷酸化状态,该调节作用可能与两者的共连接有关。     

G protein-coupled receptor kinase 4 regulates expression and phosphorylation status of AT1 receptor in rat vascular smooth muscle cells

目的 应用siRNA干扰抑制大鼠胸主动脉血管平滑肌细胞G蛋白偶联受体激酶4(G protein-coupled receptor kinase 4,GRK4)的表达,探讨GRK4对血管紧张素Ⅱ1型(angiotensinⅡtype1,AT1)受体的调节作用.方法 免疫组化检测大鼠回结肠动脉血管平滑肌组织GRK4蛋白表达;以大鼠胸主动脉平滑肌细胞株(A10细胞株)为研究对象,免疫印迹检测GRK4、AT1受体蛋白表达变化,免疫共沉淀检测GRK4和AT1受体的相互作用和AT1受体磷酸化改变.结果 大鼠动脉平滑肌组织GRK4表达良好;siRNA干扰后,GRK4蛋白表达明显下降(P＜0.05);AT1蛋白表达降低(P＜0.05),AT1受体磷酸化明显增强(P＜0.05);GRK4和AT1受体存在共连接,抑制GRK4表达后增加GRK4与AT1受体之间的共连接.结论 GRK4能够调控大鼠胸主动脉平滑肌细胞AT1受体蛋白表达及其磷酸化状态,该调节作用可能与两者的共连接有关.     

PCSK3通过调控血管平滑肌细胞炎症反应及细胞表型转化标志物的表达抑制细胞迁移

目的 ：探讨前蛋白转化酶枯草溶菌素3(PCSK3)调控小鼠血管平滑肌细胞增殖和迁移的作用。方法 ：首先采用0、25、50、100 mg/L的氧化型低密度脂蛋白（ox-LDL）刺激小鼠血管平滑肌细胞（VSMCs）,通过RT-PCR检测PCSK3 mRNA表达情况。将100 mg/L的ox-LDL刺激小鼠血管平滑肌细胞作为模型组,将VSMCs分为对照组（NC组,0 mg/L ox-LDL刺激的VSMCs+scramble siRNA,SCR）,ox-LDL组（模型组+SCR）,ox-LDL+si-PCSK3 1#组（模型组+si-PCSK3 1#质粒）以及ox-LDL+si-PCSK3 2#组（模型组+si-PCSK3 2#质粒）。EdU法检测各组VSMCs增殖活性,Transwell小室实验观察细胞迁移情况,ELISA试剂盒检测IL-1β,TNF-α,IL-6,细胞间黏附分子-1(ICAM-1)含量,RT-PCR检测各组SMCs细胞的Ⅰ型及Ⅲ型胶原,平滑肌肌动蛋白（α-SMA）的mRNA表达水平的变化。结果 ：100 mg/L ox-LDL处理浓度下的VSMCs中PCSK3明显增高,能够...     

孤儿核受体Nur77对ox-LDL诱导的血管平滑肌细胞增殖抑制作用的研究

目的探讨孤儿核受体Nur77对氧化低密度脂蛋白(ox-LDL)诱导的血管平滑肌细胞增殖的影响。方法体外原代培养大鼠血管平滑肌细胞,在ox-LDL及其相关成分7-酮胆固醇(7-KC)、9-HODE和13-HODE等条件刺激下,应用实时荧光定量反转录聚合酶链反应(RT-PCR)技术、蛋白免疫印迹技术和免疫荧光染色结合激光扫描共聚焦显微镜技术检测Nur77 mRNA和蛋白表达变化。用腺病毒感染血管平滑肌细胞过表达Nur77,通过细胞计数和Western Blotting观察Nur77对血管平滑肌细胞增殖的影响。结果 ox-LDL可诱导血管平滑肌细胞中Nur77 mRNA表达上调,并且在1 h时达到高峰;同时ox-LDL及其相关成分(7-KC、9-HODE和13-HODE),均可诱导血管平滑肌细胞中Nur77的蛋白表达明显上调。在ox-LDL刺激下,Nur77过表达组(Ad-GFP-Nur77)平滑肌细胞的细胞密度[(2.70±0.14)×104]及细胞增殖相关蛋白cyclin D1的表达明显低于对照组(Ad-GFP)平滑肌细胞的细胞密度[(3.95±0.26)×104]及cyclin D1的表达(P<0.05)。结论孤儿核受体Nur77抑制ox-LDL诱导的血管平滑肌细胞增殖。     

HDAC2 siRNA转染对人胰腺癌细胞增殖及凋亡的影响

［摘要］目的: 探讨RNA干扰沉默组蛋白去乙酰化酶2(histone deacetylase 2,HDAC2)基因表达对人胰腺癌细胞增殖和凋亡的影响。方法: 培养人胰腺癌PaTu8988细胞株,并将其随机分为5组:空白对照组,20 nmol／L HDAC2 siRNA阴性组,40 nmol／L HDAC2 siRNA阴性组,20 nmol／L HDAC2 siRNA组,40 nmol／L HDAC2 siRNA组。合成针对HDAC2基因的小干扰RNA(small interfering RNA,siRNA),利用阳离子脂质体将HDAC2 siRNA瞬时转染人胰腺癌PaTu8988细胞,分别采用实时荧光定量PCR(qRT-PCR)和蛋白质印迹法检测HDAC2基因及蛋白的表达;MTT比色法检测细胞的增殖率;流式细胞术测定细胞凋亡率。结果: 与空白对照组和阴性siRNA 组对比,HDAC2 siRNA组的HDAC2基因和蛋白的表达水平均明显下降(P均<0.05),细胞增殖率减慢(P<0.01),凋亡率明显升高(P<0.01)。结论: 采用RNA干扰沉默人胰腺癌细胞HDAC2基因表达,可抑制癌细胞增殖和诱导其凋亡。     

微小RNA干扰抑制Aurora A表达对胶质瘤细胞化疗敏感性的影响

目的:探讨微小RNA干扰抑制Aurora A表达对胶质瘤细胞化疗敏感性的影响。方法:采用Aurora A特异性siRNA对人脑胶质瘤细胞进行转染,使用Western blot和RT-PCR对Aurora A mRNA及蛋白表达情况进行检测,同时使用化疗药物尼莫司汀(ACNU)对转染后人脑胶质瘤细胞进行处理,观察并分析处理前后胶质瘤细胞的增殖和凋亡情况。结果:40 mg/L ACNU处理后各组U251细胞吸光度较0mg/L ACNU处理后各组细胞器吸光度值明显下降(P<0.05),其中以siRNA干扰组细胞吸光度值最低(0.20±0.05),细胞增殖抑制率为75.1%,远高于阴性对照组和正常对照组(P<0.05);转染Aurora A siRNA的U251细胞在经40 mg/LACNU处理72h后,其细胞凋亡率明显高于0mg/L处理组(P<0.05),且siRNA干扰组的U251细胞在培养72h后其凋亡率均明显高于正常对照组和阴性对照组,差异具有统计学意义(P<0.05)。结论:采用特异性微小RNA对Aurora A基因的表达进行干扰抑制,可有效阻碍胶质瘤细胞的增殖过程,且可有效提高胶质瘤细胞对于化疗药物的敏感性,有利于增强恶性胶质瘤术后化疗效果,对于改善患者生存质量,延长其生存时间具有十分积极的意义。     

氧化低密度脂蛋白对人脐静脉内皮细胞增殖及 circRNA-CER表达的影响

目的 探讨氧化低密度脂蛋白(ox-LDL)刺激人脐静脉内皮细胞(HUVEC)后环状RNA circRNA-CER(circRNA-100876)的表达及其对HUVEC增殖的影响。方法 分别以不同浓度ox-LDL(40、60、80、100及200 mg/L)刺激HUVEC 24 h,以及以100 mg/L ox-LDL刺激HUVEC 6、12、24及48 h,利用实时荧光定量PCR验证circRNA-CER在细胞中的表达水平;构建circRNA-CER特异性过表达质粒并进行细胞转染,采用CCK8实验研究circRNA-CER在血管内皮细胞增殖中的作用;荧光定量PCR检测circRNA-CER对miR-136表达的影响;Western blot检测过表达circRNA-CER后血管内皮生长因子(VEGF)的蛋白表达水平,荧光素酶报告基因验证circRNA-CER与miR-136、miR-136与VEGF-3′UTR的相互作用。结果 随ox-LDL处理浓度增加,circRNA-CER在HUVEC中的表达量呈下调趋势,并在100 mg/L时差异有统计学意义(P<0.05);在用该浓度ox...     

siRNA抑制THOC1基因表达对胰腺癌Panc-1细胞生物学行为的影响

目的采用RNA干扰技术下调THOC1基因在胰腺癌细胞Panc-1中的表达,探讨THOC1表达下调对Panc-1细胞生物学行为的影响,以期探讨THOC1基因的功能。方法构建靶向THOC1基因siRNA慢病毒载体(重组慢病毒中有绿色荧光蛋A真核表达框),感染Panc-1细胞(THOC1 siRNA组),并设立慢病毒载体对照组(Control-siRNA组)及空白对照组。荧光显微镜下观察并计算感染效率,Western印迹检测各组细胞中THOC1蛋白表达水平,MTT法检测细胞增殖能力,流式细胞术测定细胞周期,Transwell小室实验检测细胞体外迁移侵袭能力。结果与Control-siRNA组及空白对照组比较,THOC1 siRNA组的THOC1蛋白表达受到抑制,细胞增殖能力明显减弱(P<0.01),G1/G2期细胞数增多和S期细胞数减少表明细胞周期受阻、细胞体外迁移和侵袭能力显著减弱(P<0.01)。结论 THOC1基因具有促进胰腺癌细胞Panc-1细胞的增殖和体外迁移侵袭的作用。     

亲环素A在ox-LDL诱导的血管平滑肌细胞泡沫化过程中的作用

目的观察亲环素A在ox-LDL（氧化低密度脂蛋白）诱导血管的平滑肌细胞中的作用。方法不同浓度（0、20、40、80、160 mg/L）ox-LDL与大鼠血管平滑肌细胞共同孵育,进而选择80 mg/L ox-LDL与细胞共同孵育不同的时间（0、24、48、72、96 h）,用免疫印迹（Western blot）检测亲环素A蛋白质的表达改变。高效液相色谱法（HPLC）检测细胞内胆固醇含量。结果随ox-LDL浓度增加和孵育时间延长,亲环素A蛋白质的表达均逐渐减弱。HPLC显示80 mg/L ox-LDL孵育细胞72 h组细胞胆固醇酯/总胆固醇比值为57.9%（〉50%）,符合泡沫细胞特征。结论ox-LDL可降低血管平滑肌细胞中亲环素A的蛋白表达并诱导泡沫细胞形成。     

二烯丙基二硫对人白血病细胞增殖和钙网蛋白表达的影响

观察二烯丙基二硫(DADS)对人白血病HL-60细胞增殖和钙网蛋白(CRT)表达的影响。DADS(1.25 mg/L)处理HL-60细胞24、48和72 h,siRNA抑制CRT的表达,采用MTT法检测细胞活力,实时定量PCR和Western blot检测CRT的表达。与对照组比较,DADS处理24、48和72 h组HL-6细胞的增殖水平均显著性降低,呈现时间依赖性(均P<0.05)。与对照组比较,siRNA瞬时转染显著降低了CRT mRNA的表达(P<0.05),也显著降低了细胞的增殖水平(P<0.05),与DADS具有协同效应。DADS抑制了HL-60细胞增殖,其机制可能与下调钙网蛋白的表达有关。     

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.