艾纳香二氢黄酮对脂质过氧化损伤大鼠原代培养肝细胞的保护作用

研究 5种艾纳香二氢黄酮类化合物对过氧化损伤的原代培养肝细胞的保护作用。胶原酶灌流法分离大鼠肝细胞。以 CCl4或 Fe SO4 Cys损伤原代培养肝细胞 ,1× 10 - 4 和 1× 10 - 5 mol/ L 的 5种艾纳香二氢黄酮均能明显抑制受损伤细胞的转氨酶逸出、MDA产生及 GSH耗竭。其中以 2 ,5 -二羟基艾纳香二氢黄酮的作用最强 ,在 CCl4所致肝细胞损伤实验中 ,其抑制 MDA产生的 ED5 0 约为 2 .6× 10 - 6 mol/ L ,保护 GSH的 ED5 0 约为 1.0 8× 10 - 5 mol/ L ,抑制 AL T逸出细胞的 ED5 0 约为 9.39× 10 - 6 mol/ L     

水疱性口炎病毒对肝癌细胞的选择性感染和溶解作用

目的 :研究水疱性口炎病毒 (VSV)对肝癌细胞感染和溶解的选择性。 方法 :将 VSV分别感染 5种经过或未经过干扰素预处理的肝癌细胞系 (Hep3B、Hep G2、SMMC- 772 1、BBEL- 74 0 2和 BEL- 74 0 4细胞 )、正常传代肝细胞 (L- 0 2细胞 )和正常原代培养肝细胞 ,应用标准噬斑测定技术滴定病毒在各类细胞中的产量 ,锥虫蓝染色排除试验检测细胞活力 ,亚甲蓝和伊红染色观察细胞溶解动力学。 结果 :0 .1pfu/细胞的 VSV与上述 5种肝癌细胞、正常传代和正常原代人肝细胞共同培养 18h后 ,各细胞 VSV的产量分别为 1.8× 10 9、1.5× 10 8、8.0× 10 7、5 .6× 10 8、7.7× 10 8、6 .5× 10 7、2 .3× 10 5pfu/ m l;而经 10 0 0 U/ m l干扰素预先处理后 ,上述细胞 VSV的产量分别降至 2 .4× 10 4 、3.8× 10 4 、6 .9× 10 5、4 .2× 10 3、8.3× 10 5、5 .3× 10 4 、<10pfu/ ml。无论是否经干扰素 α- 2 b(10 0 0 U/ m l)预处理 ,除正常原代培养肝细胞外 ,VSV感染后 2 4 h其他细胞被大量溶解。结论 :VSV可迅速感染并溶解肝癌细胞及正常传代肝细胞 ,干扰素虽能完全保护正常原代肝细胞 ,但仅能部分降低 VSV在肝癌细胞和正常传代肝细胞中的增殖和溶细胞速率。说明有干扰素存在时 ,VSV能够选择性地感染和溶解肝癌细胞 ,提     

胎肝前体细胞脾移植对大鼠急性肝功能衰竭的实验研究

目的 :探讨经脾内同种异体移植培养的原代胎肝前体细胞与成体肝细胞悬液对大鼠药物性肝衰竭的疗效 ,并观察脾内移植肝细胞的生物学特性。方法 :采用 D-氨基半乳糖 ( D- gal)建立大鼠急性肝衰竭模型 ,2 4 h后随机分为三组进行治疗。 A组 :经脾内移植体外培养 7d的肝细胞 2× 1 0 7;B组 :经脾内移植肝细胞悬液 2× 1 0 7;C组 :经脾内注射生理盐水1 ml。观察受体大鼠的存活率、肝脏功能和病理变化及移植肝细胞的生物学特性。结果 :A组、B组大鼠存活率 ( 77%、5 9% )与 C组大鼠存活率 ( 2 2 % )相比具有显著性差异 ,肝功能各项指标均有明显改善 ,A、B组与 C组大鼠的肝功能改变方面有统计学差异。经 HE和 PAS染色证实 ,移植的肝细胞在受体脾内结构和功能保持较好。结论 :经脾内移植的培养的原代胎肝前体细胞和肝细胞悬液均能提高大鼠药物性肝衰竭的存活率、改善肝功能及肝脏组织病理变化 ,但培养原代胎肝前体细胞优于成体肝细胞悬液。     

链霉蛋白酶P、糖苷酶对低分子量肝细胞生长素生物活性的影响

目的研究链霉蛋白酶P、糖苷酶对低分子量肝细胞生长素生物活性的影响。方法用链霉蛋白酶P、N-糖苷酶F、O-糖苷酶分别对低分子量肝细胞生长素消化,观察消化后低分子量肝细胞生长素以及未经消化的低分子量肝细胞生长素和促肝细胞生长素对人肝癌细胞株QGY、HepG2、原代大鼠肝细胞的增殖作用,用3H-胸腺嘧啶核苷掺入法测定这些细胞的增殖情况。结果低分子量肝细胞生长素与促肝细胞生长素对人肝癌细胞株QGY、HepG2、原代大鼠肝细胞均有明显增殖作用（P〈0.01）;低分子量肝细胞生长素与促肝细胞生长素均对人肝癌细胞株QGY增殖作用最明显（P〈0.01）;低分子量肝细胞生长素增殖作用较促肝细胞生长素明显（P〈0.01）。低分子量肝细胞生长素经链霉蛋白酶P、O-糖苷酶消化前后对人肝癌细胞株QGY、HepG2、原代大鼠肝细胞增殖作用无影响（P〉0.05）;低分子量肝细胞生长素经N-糖苷酶F消化后对人肝癌细胞株QGY、HepG2、原代大鼠肝细胞增殖作用明显减弱,差异有统计学意义（P〈0.01）。结论低分子量肝细胞生长素促进人肝癌细胞株QGY、HepG2、原代大鼠肝细胞增殖作用比促肝细胞生长素更显著。     

大鼠肝细胞的分离及原代长期培养

目的 :研究大鼠肝细胞分离的简易方法及长期培养过程中肝细胞形态变化过程。方法 :采用体外胶原酶二步灌注法分离大鼠肝细胞 ,TB染色法计算细胞数及细胞活率。 MTT法观测新生牛血清对大鼠肝细胞增殖的影响。在含 1 0 %新生牛血清及其他附助因子的 Williams′E条件培养基中原代长期培养 ,并进行形态学的动态观察。结果 :平均每只大鼠可获取 2 .2 6× 1 0 8个肝细胞 ,平均活力为 95.6%。新生牛血清浓度与大鼠肝细胞增殖有明显的量效关系 (P<0 .0 1 )。在 Williams′E培养基中可存活 5～ 6周并保持正常形态。结论 :本方法分离的肝细胞有较高的获取率和活力 ,适合体外长期原代培养     

大川芎丸对血管紧张素Ⅱ受体结合活性的影响

为探讨中药复方大川芎丸(DCXW)与大鼠肝细胞膜上血管紧张素Ⅱ的Ⅰ型受体(AT1受体)的相对结合亲和力,采用蔗糖密度梯度离心法制备大鼠肝细胞膜受体制剂.通过非标记配基饱和实验建立理想的受体结合分析系统.通过竞争取代反应分别确定洛沙坦(Losartan)和DCXW的IC50值,再计算得到DCXW的相对结合亲和力.相对结合亲和力= IC50, Losartan/ IC50,DCXW×100% .通过血压测定实验确定DCXW是AT1受体激动剂还是拮抗剂.结果:Losartan的IC50值为6.5×10-3mg/ml;DCXW的IC50值为3.6×10-2mg/ml,相对结合亲和力为18.1%(相对于Losartan).DCXW有降压作用,它是AT1受体拮抗剂.以上结果表明,中药复方DCXW能够同125I AngⅡ竞争与大鼠肝细胞膜上的AT1受体结合,这与其降压和改善微循环的生理效应是一致的.     

肝纤维化大鼠星状细胞Smad3、Smad7基因表达差异的研究

通过检测肝纤维化大鼠原代肝星状细胞 (HSC)的 Sm ad3和 Smad7编码基因的表达 ,探讨 HSC活化及转化机制。皮下注射四氯化碳 (CCl4)制备大鼠肝纤维化模型 ,于注射 CCl4后 1、 4、 8周分批分离 HSC并计算 HSC平均得率 ;RT- PCR检测其 Sm ad3、Smad7m RNA的表达变化。结果显示 :1CCl4注射后第 1、 4、 8周大鼠 HSC平均得率分别为 :(3.34± 0 .36 )× 10 7/只、 (4 .2 1± 0 .6 4)× 10 7/只、 (5 .88± 0 .79)× 10 7/只 ,均高于正常大鼠 HSC平均得率(2 .2 3± 0 .19)× 10 7/只 ,组间比较有显著性差异 ,P<0 .0 5。 2与正常对照组相比 ,注射 CCl4后 1、 4、 8周大鼠 HSC中 Sm ad3m RNA表达增加 ,P<0 .0 5。注射 CCl4后 1周 Sm ad7m RNA较正常明显升高 ,P<0 .0 5 ,到第 4周和第 8周表达水平则持续下降 ,P<0 .0 1。提示 Sm ad3和 Smad7在肝纤维化发生、发展中对 HSC的活化及转化起不同的介导作用。     

大鼠肝实质及非实质细胞VLDL和HDL受体的研究

作者以~(125)I-标记的人VLDL和不含apo E的HDL_3为配体,研究了大鼠肝实质细胞(PC)及非肝实质细胞(NPC)的VLDL和HDL受体的生物学特性。结果显示:VLDL受体和HDL受体均可介导肝PC和NPC结合、摄取及降解相应的脂蛋白;肝NPC上该二受体的活性分别为PC的10倍和4倍;肝NPC上VLDL受体HDL受体的解离常数分别为15.0～34.2μg/ml和10.1～17.7μg/ml,最大结合容量分别为2170～2607ng/mg和1004～2738ng/mg细胞蛋白;VLDL受体活性受EDTA抑制及胆固醇的下降调节,HDL受体不受EDTA抑制但受胆固醇的上升调节。发现apoCⅢ对VLDL受体结合功能有明显抑制作用。     

多巴胺D_4受体对血管紧张素Ⅱ介导的血管平滑肌细胞增殖的影响

目的观察多巴胺D4受体对血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)介导的血管平滑肌细胞(vascular smoothmuscle cells,VSMCs)增殖的影响。方法以大鼠胸主动脉平滑肌细胞株(A10细胞)、原代的Wistar-Kyoto(WKY VSMCs)和自发性高血压大鼠(SHR)胸主动脉血管平滑细胞(VSMCs)为研究对象,观察刺激D4受体对AngⅡ促增殖作用的影响以及D4受体特异性阻断剂L745870对该作用的阻断效应。细胞增殖采用MTT和细胞计数方法测定。结果 AngⅡ(10-7mol/L)可促进A10细胞增殖,最大增殖幅度为64%。D4受体激动剂PD168077本身对A10细胞无增殖影响,但D4受体特异激动剂PD168077可抑制AngⅡ(10-7mol/L)介导的血管平滑肌细胞的增殖作用,最大抑制幅度为40%;应用D4受体阻断剂L745870后该抑制作用丧失;D4受体对AngⅡ介导血管增殖的作用在SHR和WKY大鼠的VSMCs之间并无区别。结论 D4受体对AngⅡ介导的血管平滑肌细胞增殖具有抑制作用,该作用可能在一定程度上抑制了高血压病所伴发的血管平滑肌细胞增殖。     

大鼠肝细胞的分离及BRL细胞培养上清对原代肝细胞增殖的影响

目的 :研究大鼠肝细胞分离的方法及正常大鼠肝细胞来源的BRL细胞培养上清对原代大鼠肝细胞贴壁和增殖的影响。方法 :采用体外胶原酶 2步灌注法分离大鼠肝细胞 ,台盼兰染色法计算细胞数及细胞活率。以四甲基偶氮唑蓝 (MTT)法检测不同浓度的BRL细胞培养上清及含体积分数 15 %胎牛血清的普通培养液对肝细胞贴壁和增殖的影响。结果 :平均每只大鼠可获 2× 10 8个肝细胞 ,平均活力 94 .3%。用普通培养液培养肝细胞 ,肝细胞贴壁率低 ;BRL细胞培养上清能明显促进肝细胞的贴壁和生长 ,且有量效关系 (P <0 .0 5 )。结论 :用本法分离的大鼠肝细胞有较高的获取率和活力 ,BRL细胞培养上清能促进正常大鼠肝细胞的贴壁及增殖     

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.