1,25二羟维生素D3对乳腺癌细胞株生长及调亡的影响

目的 研究１,２５二羟维生素Ｄ３〖１,２５（ＯＨ）２Ｄ３〗对乳腺癌细胞株ＭＣＦ－７生长及凋亡的影响。方法 采用四唑氮蓝比色（ＭＴＴ）法检测细胞增殖,光镜和电镜形态学观察,流式细胞仪测定细胞周期和凋亡率,末端脱氧核苷酸转移酶介导的原位酶标记（ＴＵＮＥＬ）法计数凋亡细胞,免疫印迹法检测ｂｃｌ－２蛋白表达。结果 １０＾－７ｍｏｌ／Ｌ的１,２５（ＯＨ）２Ｄ３就可以抑制ＭＣＦ－７增生,改变细胞周期时相分布,     

bcl-2反义寡核苷酸促进乳腺癌MCF-7细胞凋亡的研究

目的 观察ｂｃｌ－２反义寡核苷酸（ｏｌｉｇｏｎｕｃｌｅｏｔｉｄｅｓ,ＯＤＮ）对ＭＣＦ－７细胞内ｂｃｌ－２表达及细胞凋亡的影响。方法 电转染法将合成的针对ｂｃｌ－２ ｍＲＮＡ的反义ＯＤＮ导入ＭＣＦ－７乳腺癌细胞,用荧光显微镜,电镜,免疫组化法及流式细胞仪检测反义ＯＤＮ对乳腺癌细胞凋亡,ｂｃｌ－２表达及细胞周期的影响。     

抗bcl-2核酶基因转染促进GRC-1肾癌细胞凋亡的研究

目的　观察抗ｂｃｌ２ 核酶促进ＧＲＣ１ 肾癌细胞凋亡的情况。　方法　体外构建抗ｂｃｌ２核酶基因逆转录病毒真核表达载体ｐＤＯＲＲＺ,用脂质体介导的ＤＮＡ 转染法导入ＧＲＣ１ 细胞,用ＤＮＡ 和ＲＮＡ 打点杂交、流式细胞仪（ＦＣＭ） 、电镜及光镜观察。　结果　核酶（ ＲＺ） 基因成功导入ＧＲＣ１ 细胞并在其内表达,转基因后ＧＲＣ１ 细胞的ｂｃｌ２ 蛋白量明显减少,ＦＣＭ 检测转基因后ＧＲＣ１ 细胞周期Ｇ１ 期前出现了典型的亚２ 倍体凋亡峰,电镜下观察ＧＲＣ１ 细胞出现典型的凋亡现象。　结论　人工合成的抗ｂｃｌ２ 核酶基因能够成功地导入ＧＲＣ１ 肾癌细胞并抑制ｂｃｌ２ 蛋白的合成,促进ＧＲＣ１ 细胞的凋亡。     

正常成人一次性口服阿法骨化醇(1α-(OH)D_3)对骨钙素(BGP)等生化指标影响的动态观察

１α－（ＯＨ）Ｄ３在肝脏２５－羟化酶的作用下迅速转化成１，２５（ＯＨ）２Ｄ３，从而调节钙磷代谢。采用放射免疫分析方法（ＲＩＡ）对８例正常成年男性一次性口服阿法骨化醇４μｇ后，血清１，２５（ＯＨ）２Ｄ３、ＢＧＰ、Ｔ、β２－ＭＧ、ＩＧＦ－Ｉ的浓度变化进行了动态观察。结果发现，随着循环中１，２５（ＯＨ）２Ｄ３水平增加，血中ＢＧＰ、Ｔ、ＴＧＦ－Ｉ的产生显示先降后升的趋势，而β２－ＭＧ的产生则显示下降趋势     

bcl-2蛋白在肝细胞肝癌发生过程中的表达及其与肝细胞凋亡变化的关系

为探讨肝细胞肝癌发生过程中ｂｃｌ２蛋白的变化规律及与细胞凋亡变化的关系,运用ＴＵＮＥＬ法和免疫组化方法检测石蜡保存的肝细胞肝癌组织（ｈｅｐａｔｏｃｅｌｕｌａｒｃａｒｃｉｎｏｍａ,ＨＣＣ）、肝硬变组织（ｌｉｖｅｒｃｉｒｈｏｓｉｓ,ＬＣ）和正常肝组织（ｎｏｒｍａｌｌｉｖｅｒ,ＮＬ）中细胞凋亡发生率和ｂｃｌ２的表达。结果：从ＮＬ、ＬＣ到ＨＣＣ组织,细胞凋亡活性出现增强;ｂｃｌ２蛋白的表达逐渐下降,ＬＣ和ＨＣＣ组比较,Ｐ＜０．０５。结论：ｂｃｌ２的表达可能与肝细胞凋亡的抑制有关;在肝癌发生过程中,ｂｃｌ２表达下降可能是引起肝癌细胞凋亡增加的原因之一。     

bcl-2基因表达在人膀胱癌多药耐药现象中的意义

目的探讨ｂｃｌ２基因在人膀胱癌多药耐药中的作用。方法应用原位ＤＮＡ末端转移酶标记法（ＴＵＮＥＬ）、免疫细胞化学及逆转录多聚酶链式反应（ＲＴＰＣＲ）技术,分别对阿霉素诱导的人膀胱癌耐药细胞株（ＢＩＵ８７／ＡＤＭ）及敏感细胞株（ＢＩＵ８７）的细胞凋亡及其ｂｃｌ２基因表达进行研究。结果（１）在相同条件下,与敏感细胞株相比,耐药细胞株中细胞凋亡明显受到抑制;（２）耐药细胞株中ｂｃｌ２基因表达明显强于敏感细胞株。结论凋亡抑制基因ｂｃｌ２在人膀胱癌多药耐药细胞中的高表达,是导致其细胞凋亡受抑制的重要原因,对于人膀胱癌多药耐药的产生具有重要意义。     

肝门部胆管癌组织内血管形成与细胞凋亡相关性的研究

目的　研究肝门部胆管癌中细胞凋亡和血管形成的相互关系。方法　采用原位末端标记技术和抗因子Ⅷ的抗体分别检测肝门部胆管癌细胞凋亡指数和瘤内微血管的密度,同时用免疫组化法检测血管内皮细胞生长因子（ＶＥＧＦ） 和胆管癌细胞凋亡调控基因ｂｃｌ２ 的表达。结果　肝门部胆管癌组织ＶＥＧＦ阳性表达率５５－５６％ （２０／３６）,ＶＥＧＦ阳性表达与ＭＶＤ呈正相关（Ｐ＜ ０－０１） 。高血管组的ｂｃｌ２ 阳性表达明显高于低血管组（ Ｐ＜ ０－０５）,而低血管组的细胞凋亡指数高于高血管组（ Ｐ＜０－０１）。ｂｃｌ２ 表达丰富的区域,ＶＥＧＦ的表达率高。结论　肝门部胆管癌血管形成影响细胞凋亡的敏感性和发生率。     

大肠癌组织酶标志物研究

目的：探讨大肠癌的发病机理及早期诊断的酶标志物。方法：采用酶学方法测定大肠癌及近心端和远心端癌旁组织中鸟氨酸脱羧酶（ＯＤＣ）、γ－谷氨酰转肽酶（γ－ＧＴ）、乳酸脱氢酶（ＬＤＨ）活性及ＬＤＨ同工酶谱。结果：（１）大肠癌组织中ＯＤＣ、γ－ＧＴ及ＬＤＨ活性均显著高于癌旁正常组织；（２）大肠癌组织中ＬＤＨ５同工酶及ＬＤＨ４＋ＬＤＨ５／ＬＤＨ１＋ＬＤＨ２的比值也显著升高，ＬＤＨ同工酶谱向Ｍ型转移。结论：肠组织中ＯＤＣ、γ－ＧＴ、ＬＤＨ活性升高，ＬＤＨ同工酶谱向Ｍ型转移与大肠癌的形成有关，提示ＯＤＣ、γ－ＧＴ及ＬＤＨ可能是大肠癌的组织酶标志物     

Rxa对L02细胞增殖的细胞周期分析和对Ki-67、CyclinD1、PCNA蛋白表达的影响

目的　探讨丹参提取物Ｒｘａ 对正常肝细胞（Ｌ０２ 细胞） 增殖速率的影响。方法　应用流式细胞术（ＦＣＭ）分别测定Ｌ０２ 细胞Ｒｘａ 处理组（Ｌａ 组） 和未处理组（Ｌ组）０、２、４、８、１２ 小时Ｇ０Ｇ１ 、Ｓ、Ｇ２Ｍ 期细胞指数,同步进行Ｋｉ６７、Ｃｙｃｌｉｎ Ｄ１ 、ＰＣＮＡ 蛋白表达的免疫组化分析。结果　Ｌａ 组细胞增殖速率较快,Ｋｉ６７ 抗原表达相对较强,ＣｙｃｌｉｎＤ１、ＰＣＮＡ 表达相对提前。结论　Ｒｘａ 可能促进Ｌ０２ 细胞增殖。     

1α，25双羟维生素D3对成骨样细胞增殖与分化的影响

采用同位素掺入，细胞周期、细胞化学和扫描电镜等方法观察了１α，２５双羟维生素Ｄ３［１，２５（ＯＨ）２Ｄ３］对人及大鼠成骨样细胞ＯＳ－７３２和ＲＯＳ１７／２．８增殖及分化的影响。结果表明：１，２５（ＯＨ）２Ｄ３对ＯＳ－７３２细胞增殖的抑制作用呈明显的时效和量效关系。在给１０－７ｍｏｌ／Ｌ的１，２５（ＯＨ）２Ｄ３后第４和第６天，对ＯＳ－７３２细胞生长的抑制率分别为４０％和６０％；对ＤＮＡ，ＲＮＡ和蛋白质合成的抑制作用分别为５９％，４１％和２２％。流式细胞计测定结果表明：１，２５（ＯＨ）２Ｄ３使ＤＮＡ合成受阻；扫描电镜显示：１，２５（ＯＨ）２Ｄ３有抑制ＲＯＳ１７／２．８细胞表面微绒毛的作用。此外，细胞化学染色表明：该激素有增加成骨样细胞碱性磷酸酶活性和促进骨形态形成蛋白合成的作用，即刺激骨形成的作用。     

Vasopressor hormone response following mesenteric traction during major abdominal surgery

Background : We investigated the vasopressor hormone response following mesenteric traction (MT) with hypotension due to prostacyclin (PGI₂) release in patients undergoing abdominal surgery with a combined general and epidural anesthesia. Methods : In a prospective, randomized, placebo-controlled study we administered 400 mg ibuprofen (i.v.) in 42 patients scheduled for abdominal surgery. General anesthesia was combined with epidural anesthesia (T4-L1). Before as well as 5, 15, 30, 45, and 90 min after MT we recorded plasma osmolality, hemodynamics and measured 6-keto-PGFlα (stabile metabolite of PGI₂), TXB₂ (stabile metabolite of thromboxane A₂) active renin, and arginine vasopressin (AVP) plasma concentrations by radioimmunoassay. Catecholamine levels were assessed by high-pressure liquid chromatography (HPLC) with electrochemical detection. Results : Following MT, arterial hypotension occurred along with a substantial PGI₂ release. This was completely abolished by ibuprofen administration. Although plasma levels of 6-keto-PGF_1α (1133 (708) vs. 60 (3) ng/L, median (median absolute deviation), P=0.0001, placebo vs. ibuprofen) remained significantly elevated, blood pressure was restored within 30 min after MT in the placebo group. At the same point in time plasma concentrations of TXB² (164 (87) vs. 58 (1) ng/L, P=0.0001), epinephrine (46 (33) vs. 14 (6) ng/L, P=0.001), AVP (41 ± (18) vs. 12 (7) ng/L, P=0.0004), and active renin (27 (12) vs. 12 (4) ng/L, P = 0.001) were significantly higher in placebo-treated patients. Conclusion : Under combined general and epidural anesthesia arterial hypotension following MT due to endogenous PGI₂ release is associated with enhanced release of AVP, active renin, epinephrine and thromboxane A₂, presumably contributing to hemodynamic stability within 30 min after MT.     

A Teaspoon Pump for Pumping Blood with High Hydraulic Efficiency and Low Hemolysis Potential

Abstract: Virtually all blood pumps contain some kind of rubbing, sliding, closely moving machinery surfaces that are exposed to the blood being pumped. These valves, internal bearings, magnetic bearing position sensors, and shaft seals cause most of the problems with blood pumps. The original teaspoon pump design prevented the rubbing, sliding machinery surfaces from contacting the blood. However, the hydraulic efficiency was low because the blood was able to "slip around" the rotating impeller so that the blood itself never rotated fast enough to develop adequate pressure. An improved teaspoon blood pump has been designed and tested and has shown acceptable hydraulic performance and low hemolysis potential. The new pump uses a nonrotating "swinging" hose as the pump impeller. The fluid enters the pump through the center of the swinging hose; therefore, there can be no fluid slip between the revolving blood and the revolving impeller. The new pump uses an impeller that is comparable to a flexible garden hose. If the free end of the hose were swung around in a circle like half of a jump rope, the fluid inside the hose would rotate and develop pressure even though the hose impeller itself did not "rotate"; therefore, no rotating shaft seal or internal bearings are required.     

Microspheres Based Detoxification System: A New Method in Convective Blood Purification

Abstract: A variety of protein-bound or hydrophobic substances, accumulating as a result of pathologic conditions such as exogenous or endogenous intoxications, are removed poorly by conventional detoxification methods because of low accessibility (hemodialysis), insufficient adsorption capabilities (hemosorption), low efficiency (peritoneal dialysis), or economic limitations (high-volume plasmapheresis). Combining advantages of existing methods with microspheric technology, a module-based system was designed. Major operating parameters of the latter can be modified to allow for adjustment to individual clinical situations. An extracorporeal blood circuit including a plasmafilter is combined with a secondary high-velocity plasma circuit driven by a centrifugal pump. Different microspheric adsorbers can be combined in one circuit or applied in sequence. Thus, a prolonged treatment can be tailored using specially designed selective adsorber materials. Comparing this system with existing methods (high-flux hemodialysis, molecular adsorbent recycling system), results from our in vitro studies and animal experiments demonstrate the superior efficiency of substance removal.     

Tissue perfusion in relation to cardiac output during continuous positive-pressure ventilation and administration of propranolol or verapamil

Background : Our objective was to determine whether administration of propranolol or verapamil modifies the hemodynamic adaptation to continuous positive-pressure ventilation (CPPV), in particular the regional distribution of cardiac output (CO).
Methods : General hemodynamics and regional blood flows assessed by microsphere technique (15 (μm) were recorded in 16 anesthetized pigs during spontaneous breathing (SB) and CPPV with 8 cm H₂O end-expiratory pressure (CPPV8) before and after intravenous administration of propranolol (0.3 mg · kg⁻¹ followed by 0.15 mg · kg⁻¹ · h⁻¹, n=8) or verapamil (0.1 mg · kg⁻¹ followed by 0.3 mg · kg⁻¹ · h⁻¹, n=8).
Results : CPPV8 depressed CO by 25% without shifts in its relative distribution with the exception of a noteworthy increase in adrenal perfusion. Propranolol increased arterial blood pressure, and due to a fall in heart rate, CO dropped by 25%. The kidneys and, to a lesser extent, the splanchic region and central nervous system received increased fractions of the remaining CO at the expense of skeletal muscle flow. Similar patterns were seen during SB and CPPV8 such that the combination of propranolol and CPPV8 depressed CO by 50%. The circulatory effects of verapamil were less evident but myocardial perfusion tended to increase.
Conclusions : The combination of propranolol or verapamil with CPPV does not result in any specific hemodynamic interaction in anesthetized pigs, except that the combined effect of propranolol and CPPV may severely reduce CO.     

Bariatric Surgery in Some "Central and East-European (former Eastern bloc)"Countries - Current Status and Prediction for the Next Millennium

Background: Obesity is increasing globallly, including in the formerly "Eastern Bloc" countries. Methods: A survey was made of obesity and bariatric surgery. Results: In the 8 East and Central European countries studied, with total population 300 million, roughly 43% of the population was overweight (BMI 25-30), 23% obese (BMI > 30), with about 15 million people morbidly obese (BMI > 40). From 0-10 morbidly obese individuals/100,000/year undergo bariatric surgery. Conclusion: Most countries were found to provide inadequate treatment for obesity.The majority of the morbidly obese are not treated effectively. However, health-care awareness of obesity and bariatric surgeons are slowly increasing.     

Volatile anaesthetics reduce adhesion of blood platelets under low-flow conditions in the coronary system of isolated guinea pig hearts

Background : Inhibitory effects of volatile anaesthetics on platelet aggregation have been demonstrated in several studies. However, the influence of volatile anaesthetics on intracoronary platelet adhesion has not been elucidated so far.
Methods : Isolated hearts of guinea pigs were perfused with buffer in the absence or presence of volatile anaesthetics (0.5 and 1 MAC) at constant coronary flow rates of 5 ml/min for 25 min, then 1 ml/min for 30 min and again 5 ml/min for 10 min. Before, during and after low-flow perfusion, a bolus of human platelets was applied into the coronary system. To simulate thrombogenic conditions, 0.3 U/ml human thrombin was infused during low-flow perfusion and reperfusion. The number of platelets sequestered to the endothelium was calculated from the difference between coronary in- and output of platelets. The myocardial production of lactate and consumption of pyruvate and coronary perfusion pressure were also determined.
Results : At a flow rate of 5 ml/min only about 3% of the applied platelets did not emerge from the coronary system, in any group. In contrast, 13.1±1.2% (mean±SEM) of infused platelets became adherent in low-flow perfusion in the control group without anaesthetic. The adherence was reduced with each 1 MAC isoflurane (to 6.2±1.2%), sevoflurane (to 4.4±0.9%) or halothane (to 3.2±1.5%) (each P <0.05 vs. control). Volatile anaesthetic, 0.5 MAC, did not inhibit platelet adhesion to a statistically significant extent in any case. Perfusion pressure and metabolic parameters were not statistically different between the control and the hearts exposed to anaesthetics.
Conclusion : Volatile anaesthetics in a concentration of 1 MAC can reduce the adhesion of platelets in the coronary system under reduced flow conditions. This action does not arise from vasodilation or inhibition of ischaemic stress.     

Synergistic interaction between the effects of propofol and midazolam with fentanyl on phrenic nerve activity in rabbits

Background: It has been shown that the depressive effects of both propofol and midazolam on consciousness are synergistic with opioids, but the nature of their interactions on other physiological systems, e. g. respiration, has not been fully investigated. The present study examined the effect of propofol and midazolam alone and in combination with fentanyl on phrenic nerve activity (PNA) and whether such interactions are additive or synergistic. Methods: PNA was recorded in 27 anaesthetised and artificially ventilated rabbits. In three groups, propofol, fentanyl and midazolam were administered intravenously in incremental doses to construct dose-response curves for the depressant effects of each one on PNA. In another two groups, the effect of pretreatment with either fentanyl 1 μg · kg^?1 i. v. or midazolam 0.05 mg · kg^?1 i. v. on the effects of propofol and fentanyl respectively on PNA were studied. Results: Propofol and fentanyl caused a dose-dependent depression of PNA with complete abolition at the highest total doses of 16 mg · kg^?1 i. v. and 32 μg · kg^?1 i. v., respectively. In contrast, midazolam in incremental doses to a total of 0.8 mg · kg^?1 reduced mean PNA by 63%, but approximately 12% of PNA remained at a total dose as high as 6.4 mg · kg^?1. The mean ED₅₀s, calculated from dose-response curves, were 5.4 mg · kg^?1, 3.9 μg · kg^?1 and 0.4 mg · kg^?1 for propofol, fentanyl and midazolam, respectively. Initial doses of either fentanyl 1 μg · kg^?1 i. v. or midazolam 0.05 mg · kg^?1 i. v. acted synergistically with subsequent doses of either propofol or fentanyl to abolish PNA at total doses of 8 mg · kg^?1 and 8 μg · kg^?1, respectively. Conclusion: Fentanyl has a synergistic interaction with both propofol and midazolam on PNA and hence potentially on respiration.     

Influence of dopexamine hydrochloride on haemodynamics and regulators of circulation in patients undergoing major abdominal surgery

Background: Catecholaminergic support is often used to improve haemodynamics in patients undergoing major abdominal surgery. Dopexamine is a synthetic vasoactive catecholamine with beneficial microcirculatory properties. Methods: The influence of perioperative administration of dopexamine on cardiorespiratory data and important regulators of macro- and microcirculation were studied in 30 patients undergoing Whipple pancreaticduodenectomy. The patients received randomized and blinded either 2 μg · kg^?1 · min^?1 of dopexamine (n=15) or placebo (n=15, control group). The infusion was started after induction of anaesthesia and continued until the morning of the first postoperative day. Endothelin-1 (ET-1), vasopressin, atrial natriuretic peptide (ANP), and catecholamine plasma levels were measured from arterial blood samples. Measurements were carried out after induction of anaesthesia, 2 h after onset of surgery, at the end of surgery, 2 h after surgery, and on the morning of the first postoperative day. Results: Cardiac index (CI) increased significantly in the dopexamine group (from 2.61±0.41 to 4.57±0.78 1 · min^?1 · m^?2) and remained elevated until the morning of the first postoperative day. Oxygen delivery index (DO₂I) and oxygen consumption index (VO₂I) were also significantly increased in the dopexamine group (DO₂I: from 416±91 to 717±110 ml/m² · m²; VO₂I: from 98±25 to 157±22 ml/m² · m²), being significantly higher than in the control group. pHi remained stable only in the dopexamine patients, indicating adequate splanchnic perfusion. Vasopressive regulators of circulation increased significantly only in the untreated control patients (vasopressin: from 4.37±1.1 to 35.9±12.1 pg/ml; ET-1: from 2.88±0.91 to 6.91±1.20 pg/ml). Conclusion: Patients undergoing major abdominal surgery may profit from prophylactic perioperative administration of dopexamine hydrochloride in the form of improved haemodynamics and oxygenation as well as beneficial influence on important regulators of organ blood flow.     

Immunomodulation in Transplantation with Photopheresis

Abstract: Photopheresis is a technique in which peripheral blood mononuclear cells, in the presence of a photoacti-vatable compound, are exposed extracorporeally to ultraviolet A light and reinfused, inducing a host autoregula-tory immune response. Experimental work and ongoing clinical studies are helping to define the role of this novel, safe, and non-toxic immunomodulating technology in the field of transplantation.