9种不同心血管风险的临床药物对hERG钾通道的作用

目的：在GLP实验室评估膜片钳检测临床用药物对hERG（human ether?à?go?go related gene）钾通道作用的差异性和重复性，研究9种致尖端扭转型室速（TdP）风险的临床用药物（高风险临床用药物：苄普地尔、奎尼丁、索他洛尔；中风险临床用药物：昂丹司琼、西沙比利、特非那定；低风险临床用药物：雷诺嗪、维拉帕米和美西律）对hERG钾通道的阻断作用。方法：采用全细胞膜片钳技术记录不同浓度的苄普地尔、奎尼丁、索他洛尔、昂丹司琼、西沙比利、特非那定、雷诺嗪、维拉帕米和美西律作用于外源稳定转染表达hERG钾通道的HEK293细胞（hERG?HEK293稳态细胞）后hERG电流（IKr）的变化，研究上述临床用药物对IKr作用的浓度依赖性及半数抑制浓度（IC50）。结果：9种临床用药物对hERG?HEK293细胞上IKr作用具有浓度依赖性，且高风险临床用药物苄普地尔和奎尼丁的IC50值分别为98.32 nmol/L和1.95 μmol/L，索他洛尔的IC50值大于300 μmol/L；中风险临床用药物昂丹司琼、西沙比利和特非那定的IC50值分别为0.94 μmol/L、39.10 nmol/L和128.58 nmol/L；低风险临床用药物雷诺嗪、维拉帕米和美西律的IC50值分别为9.94 μmol/L、235.49 nmol/L和65.56 μmol/L。本实验所得临床用药物IC50值基本与文献相符。结论：临床用药物致TdP风险与hERG通道的阻滞作用密切相关，但hERG通道阻断不等同于TdP风险，还与心脏上表达的多种离子通道有关，某些临床用药物可以通过阻断钠通道和钙通道而降低风险。本研究结果提示本方法所得数据可靠，为国内GLP实验室进行hERG钾通道评价研究提供了参考依据，可用于药物心脏毒性评价。     

传统抗心律失常药物及苦参碱对家兔心室肌细胞快速延迟整流钾电流作用的比较

目的 比较传统的抗心律失常药物和苦参碱时心肌细胞快速延迟整流钾电流(IKr)的作用,以及其诱发QT间期延长和致心律失常作用发生的危险性.方法 采用酶解法急性分离单个家兔心室肌细胞,全细胞 膜片钳技术观察并比较奎尼丁、普萘洛尔、胺碘酮及苦参碱等药物对心肌细胞IKr的作用.结果 胺碘酮(0.1、1和10μM)及苦参碱(1、10和100μM)剂量依赖性抑制家兔心室肌细胞IKr,而苦参碱的抑制作用明显弱于胺碘酮;几种传统的抗心律失常药物中,在相同药物浓度下(1μM),奎尼丁和胺碘酮均表现为明显的IKr电流抑制作用,而普萘洛尔对IKr电流的抑制作用明显减轻(n=12,P<0.05).结论 中药苦参碱及β受体阻断药普荼洛尔对IKr电流的抑制作用较奎尼丁和胺碘酮为弱,其临床应用不易诱发QT间期延长和心律失常.     

多非利特与维拉帕米联合应用对豚鼠乳头肌动作电位的影响

目的 研究钾通道拮抗剂多非利特与钙通道拮抗剂维拉帕米联合应用对豚鼠乳头肌动作电位各参数的影响.方法 应用标准微电极技术记录离体豚鼠乳头肌动作电位,观察在不同刺激频率下(0.2,0.5 ,1.0,1 25,2.0 Hz),不同浓度的胺碘酮(1,5,10 μmol/L)、多非利特(10,50,100 nmol/L)或100 nmol/L多非利特与1 μmol/L 维拉帕米联合应用对乳头肌动作电位的影响.结果 胺碘酮浓度依赖性的延长动作电位复极50%(APD50)和90%(APD90)的时程,其作用不表现为频率依赖性;多非利特延长APD具有明显的浓度依赖性,刺激频率增加时,延长APD的作用明显减弱,表现为明显的负性频率依赖性;100 nmol/L多非利特与1 μmol/L维拉帕米联用后,上述频率依赖性得到改善.结论 胺碘酮和多非利特均能浓度依赖性地显著延长动作电位时程,但复合性Ⅲ类抗心律失常药物胺碘酮对频率的依赖性明显低于纯Ⅲ类抗心律失常药物多非利特,而多非利特与维拉帕米联合应用达到一定的平衡状态时可以减少这种负性频率依赖性作用.     

藤黄酸对 K562 细胞 hERG 钾通道蛋白的调控作用

目的 探讨藤黄酸对白血病细胞系 K562 增殖、凋亡、细胞周期的影响,并观察藤黄酸对 hERG 钾通道蛋白的调控作用。方法 K562 细胞以不同浓度藤黄酸 (0.125～8.0 μmol/L) 处理 0～72 h 后,MTT 法观察藤黄酸对 K562 细胞生长抑制的情况,Annexin-V/PI 双标法及透射电镜检测细胞凋亡,PI 单染法检测细胞周期分布,Western blotting、RT-PCR 法分别检测藤黄酸对 K562 细胞内 hERG 通道的调控作用。结果 藤黄酸能明显抑制 K562 细胞增殖,具有时间-剂量依赖性,其 24 h 的 IC₅₀ 为 (2.637±0.208) μmol/L。此外,藤黄酸以浓度依赖性方式诱导 K562 细胞凋亡,并伴随明显的凋亡细胞形态学改变,而藤黄酸的凋亡诱导效应可能与其诱导 K562 细胞周期阻滞于 G₀/G₁ 期有关。hERG 钾通道蛋白在人白血病 K562 细胞中表达量较高,藤黄酸对 hERG 钾通道蛋白及其表达水平均有不同程度的抑制作用,该抑制作用呈明显的量效关系 (P＜0.01)。结论 藤黄酸可通过下调 hERG 钾通道蛋白的表达发挥较强的抗白血病效应,hERG 钾通道有望成为白血病诊治的新靶标。     

人类果蝇相关基因(HERG)通道与药物相关性长QT综合征的关系研究

人类果蝇相关基因一HERG(human ether-go-gorelated gene)基因编码心脏快速延迟整流钾通道(IKr)的α亚基[1],快速延迟整流钾电流在心肌复极化过程中起着重要作用,当HERG钾离子通道被药物阻断时,将会延长动作电位时程(APD)及QT间期,导致长QT综合症(LQTS)并诱发尖端扭转型室性心动过速(TdP)等快速型室性心律失常的发生.     

抗心律失常药物的临床合理应用

目的 了解心律失常患者的临床用药情况,并分析其合理性.方法 采用回顾性调查方法,在医院病历管理系统随机 查阅2007年4月～2008年4月间心律失常患者住院病例355份(入选300份),对临床药物利用、联合用药情况及临床应用的合理性进行统计、分析、得出结论.结果 在用药物品种上,I类钠通道阻滞剂、β-受体阻滞剂、钙通道阻滞剂,常用的代表药物为奎尼丁、普鲁卡因胺、利多卡因、美西律、普罗帕酮、氟卡尼、普萘洛尔、索他洛尔、胺碘酮、溴苄胺、维拉帕米,严格根据患者的临床症状选择药物,部位联合用药.结论 该院抗心律失常药物应用基本合理,与国际用药趋势一致.     

应用hiPSC⁃ACM和hiPSC⁃VCM进行药物毒性检测的研究

目的：建立可用于评价药物心脏毒性的人源性诱导多能干细胞分化的心房、心室肌细胞（human induced pluripotent stem cell derived atrial and ventricular cardiomyocytes,hiPSC?ACM、hiPSC?VCM）模型,并采用此模型评价化合物潜在的心脏毒性。方法：通过调控视黄酸（retinoic acid,RA）通路分化出目标细胞,分为RA处理组（RA）和RA抑制剂处理组（RAi）,用流式细胞法以及免疫荧光染色法检测心室特异性标志物（MLC2v）和心肌细胞特异性标志物（α?actinin）,荧光定量RT?PCR法检测细胞心房、心室基因（MLC2v、MYH7、NR2F2、KCNA5）表达情况,通过细胞活性实验（CCK8）检测已知致心律失常药物特非那定、索他洛尔、伊布利特在不同浓度（0.1、1.0、100.0、1 000.0 μmol/L）下对细胞的活性影响,通过钙敏染料Fluo?3 AM检测特非那定（50、100、500 nmol/L）、索他洛尔（1、10、100 μmol/L）、伊布利特（0.1、1.0、10.0 μmol/L）对于细胞钙瞬变的影响。结果：流式、免疫荧光染色以及荧光定量RT?PCR均显示,心房肌细胞特异性标志物在RA组高表达而在RAi组低表达,心室肌细胞标志物在RA组低表达而在RAi组高表达。RA可以促进hiPSCs向心房肌（ACM）分化,RAi可以促进hiPSCs向心室肌（VCM）分化。特非那定浓度达到较高浓度（100 μmol/L）时,两组细胞的活性显著下降,而两组细胞钙瞬变振幅在较低浓度（50 nmol/L）时即出现显著下降。在高浓度索他洛尔（1 mmol/L）的刺激下两组细胞活性显著下降,同样伊布利特的浓度达到1 mmol/L时两组细胞活性均有显著下降。hiPSC?VCM在不同浓度索他洛尔以及伊布利特的刺激下钙瞬变振幅均无显著变化,然而hiPSC?ACM在特非那定1 μmol/L及索他洛尔1 μmol/L刺激下即开始出现振幅显著下降。结论：人源性诱导多能干细胞可以通过调控视黄酸通道分化的心房、心室肌细胞建立体外心脏毒性评价模型,通过检测其钙瞬变的改变可以迅速、准确地评价药物毒性。     

粉防己碱对豚鼠心肌细胞钾通道的影响

目的 :研究粉防己碱对豚鼠心肌细胞钾通道的影响。方法 :用内面向外膜片钳单通道记录法。结果 :Tet10 μmol·L-1时对钾通道影响不大 (P >0 .0 5 ) ,2 0 ,30 μmol·L-1使开放概率分别由药前的 0 .775± 0 .35 6降至 0 .2 31± 0 .2 73和 0 .2 2 1± 0 .2 30 ;变化幅度 (% )为 - 6 9.13± 2 9.6和 - 74 .93± 2 3.6 3(P <0 .0 5 )。结论 :一定浓度下Tet对心肌细胞钾通道具有阻滞作用。     

不同浓度藏红花酸对HERG钾通道蛋白表达的影响

目的 了解不同浓度藏红花酸对HERG钾通道蛋白的影响.方法 采用Western blotting法、免疫荧光化学染色法和激光共聚焦显微镜检测法,检测不同浓度藏红花酸对稳定转染有HERG通道的HEK-293细胞中HERG钾通道蛋白表达的影响.结果 1、3、10和30 μmol·L-1藏红花酸对HERG钾通道蛋白的表达均无明显影响.结论 藏红花酸对HERG钾通道蛋白的表达无明显影响,作为抗心律失常药物成分具有较好的安全性.     

比那地尔与肺动脉平滑肌细胞的增殖和凋亡

目的:研究钾通道开放剂比那地尔(Pin)对大鼠肺动脉平滑肌细胞(PASMC)增殖和凋亡的作用.方法:通过非核素标记细胞增殖检测、3H-TdR掺入、形态学、流式细胞术等方法研究Pin对大鼠PASMC增殖和凋亡的作用.结果:25～500 μmol/L的Pin呈剂量依赖性地减少大鼠PASMC的细胞数和DNA合成,500 μmol/L的Pin显著减少S期和G2/M期的细胞比例,增加G0/G1期细胞的比例.50～500 μmol/L的Pin呈剂量依赖性地增加大鼠PASMC的凋亡率.结论:钾通道开放剂抑制大鼠PASMC增殖并促进其凋亡.钾通道开放剂可能成为今后预防和控制低氧性肺动脉高压血管重建的有效药物.     

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.