半定量检测胆管癌组织中人fxyd6基因的表达

目的研究人fxyd6基因在正常胆管与胆管癌组织中的表达，检测该基因在胆管癌组织切片中的亚细胞表达定位。方法采用逆转录-聚合酶链反应（RT-PCR）方法检测人fxyd6基因在正常胆管和胆管癌新鲜组织中的表达；采用原位RT-PCR（IS-RT-PCR）方法原位检测该基因在胆管癌石蜡组织切片中的亚细胞表达定位，并应用多媒体图像分析软件进行半定量分析。结果人fxyd6基因在正常胆管和胆管癌组织中存在差异性表达；原位RT-PCR检测表明人fxyd6基因定位于胆管癌上皮细胞胞浆内，在肿瘤细胞中的表达明显高于在正常胆管细胞中的表达（P〈0．05）。结论人fxyd6基因在胆管癌的转化、演进过程中可能起重要作用。     

肝外胆管癌组织BCL-2中的表达及意义

目的:了解抗调亡基因BCL-2在肝外胆管癌和良性胆管疾病组织中的表达及其与胆管癌分化程度的关系。方法:利用免疫组织化学方法检测了46例肝外胆管癌,10例胆总管囊肿,5例慢性胆管炎胆管壁组织BCL-2蛋白的表达。结果:胆管癌BCL-2蛋白表达阳性率为(43.3％)。其中高分化腺癌BCL-2蛋白阳性率为(64.3％),中分化腺癌BCL-2蛋白阳性率为(42.8％),低分化腺癌BCL-2蛋白阳性率(18.1％)。10例胆总管囊肿及5例慢性胆管炎的胆管壁组织均无表达。结论:1.BCL-2蛋白在胆管癌与良性胆管疾病中表达有显著差异,它的过度表达与肝外胆管癌的发生有关。2.胆管癌分化程度越高,其BCL-2表达越高,而分化程度越低,其BCL-2表达越低。提示BCL-2蛋白在肝外胆管癌表达与细胞分化程度有关。     

Survivin基因在胆管癌组织中的表达及其与bcl-2和bax相关性

目的 探讨Survivin基因在胆管癌组织中的表达以及与bel-2和bax基因表达的相关性.方法 应用逆转录-聚合酶链反应(RT-PCR)法检测Survivin mRNA在64例胆管癌,22例癌旁组织及10例正常胆管组织中的表达,免疫组织化学方法测定胆管癌组织中bcl-2和bax蛋白的表达.结果 胆管癌组织中Survivin基因阳性表达率65.6%(42/64),显著高于其在癌旁组织(9.1%,P<0.01)和正常胆管(0%,P<0.01)中的表达;Survivin基因表达与胆管癌的性别、年龄、部位、分化程度、临床分期及淋巴结转移无明显相关(P>0.05).bcl-2和bax在胆管癌组织中的表达率分别为56.3%(36/64)N45.3%(29/64),Survivin基因表达与bcl-2表达正相关(r=0.404,P<0.01),与bax表达负相关(r=-0.373,P<0.01).结论 Survivin基因在胆管癌的发生、发展过程中可能起重要作用;Survivin有可能成为胆管癌基因治疗的新靶点;在胆管癌的发展中,Survivin与bel-2和bax分别起协同和拮抗作用.     

Survivin在胆管癌中的表达

目的　探讨Survivin基因表达与胆管癌发生发展的关系。方法　应用免疫组织化学技术 (SP法 )检测Survivin基因在 3 3例胆管癌标本、2 8例癌旁胆管组织和 5例胆管良性病变标本中的表达。结果　 3 3例胆管癌组织中Survivin阳性表达率为 72 .7% ( 2 4/3 3 ) ;在癌旁胆管组织和胆管良性病变组织中未检测到Survivin的表达 ,胆管癌组织、癌旁胆管组织、胆管良性病变组织之间的Survivin基因的表达差异有非常显著性 (P <0 .0 1)。Survivin基因的表达与患者的性别、年龄、肿瘤的大小、分化及是否转移无关。结论　Survivin在胆管癌中高度表达可能与胆管癌的发生发展有关 ,与预后的关系有待进一步的探讨。     

核酸内切酶在肝门部胆管癌组织和细胞株中的表达和临床意义

目的 研究核酸内切酶(Dicer)在肝门部胆管癌组织和胆管癌细胞株(QBC939)中的表达及其与临床病理因素和预后的相关性.方法 采用免疫组化染色检测40例肝门部胆管癌和10例正常胆管组织中Dicer的表达;Western-blot和RT-PCR检测人胆管癌细胞株QBC939与人胆管上皮细胞株HIBEpic中Dicer的表达量;分析Dicer表达与临床病理因素的相关性.对根治性切除的肝门部胆管癌患者术后总生存期和无病生存期进行单因素和多因素分析.结果 Dicer在肝门部胆管癌组织中的表达明显低于正常胆管组织(P＜0.05),在QBC939中的表达明显低于HIBEpic(P＜0.05),高分化胆管腺癌中Dicer表达明显高于中、低分化腺癌.单因素生存分析结果显示Dicer低表达组的根治性切除术后总生存期和无病生存期均短于高表达组(P＜0.01).多因素分析显示Dicer表达水平为与预后相关的独立危险因素(P＜0.05).结论 在肝门部胆管癌和胆管癌细胞株Dicer表达下调;低表达Dicer的胆管癌患者预后更差.Dicer可能为肝门部胆管癌的诊断和判断预后的分子标志物.     

Bcl-2和Bax在胆管癌中的表达及意义

目的　检测 4 9例肝外胆管癌和 10例正常胆管Bcl 2和Bax的表达。方法　免疫组化ABC法。结果　Bcl 2在胆管癌中阳性表达 (48.98% )显著高于正常胆管 (10 .0 0 % ) (P <0 .0 5 ) ;高、中分化胆管癌Bcl 2阳性表达显著高于低分化癌 (P <0 .0 5 ) ;胆管癌Bax阳性表达 (5 3.0 6 % )与正常胆管 (80 .0 0 % )相比无统计学意义 (P >0 .0 5 ) ,正常胆管Bax阳性表达显著高于Bcl 2阳性表达 (P<0 .0 1) ,而胆管癌中Bcl 2与Bax阳性表达无显著性差异 (P >0 .0 5 )。结论　Bcl 2过度表达对胆管癌的发生可能起促进作用 ;胆管癌中Bcl 2低表达可能是预后不良的预测指标之一。     

增殖细胞核抗原蛋白和mRNA在胆管癌中的表达及其意义

目的 检测胆管癌和癌旁0.5cm胆管组织及手术切缘的正常胆管组织增殖细胞核抗原(PCNA)蛋白及mRNA的表达,探讨其在胆管癌发生发展中的作用.方法 应用免疫组织化学过氧化物酶标记链霉卵白素法(SP)检测42例胆管癌组织及20例正常组织中PCNA蛋白的表达,同时应用逆转录-聚合酶链反应(RT-PCR)技术检测42例术中所取的新鲜的胆管癌、17例同个体癌旁胆管黏膜和20例正常胆管组织中的PCNA mRNA表达,并与临床资料进行相关分析.结果 正常胆管组织、癌旁组织、胆管癌组织中PCNA mRNA相对表达量分别0.5605±0.0331,0.5692±0.0408,0.6303±0.0773,PCNA mRNA在3组之间表达呈上升趋势(P<0.05),PCNA蛋白表达趋势同其相对应的基因表达趋势一致,即PCNA mRNA在胆管癌组织中高表达(P<0.05).结论 PC-NA基因转录和蛋白可能参与了胆管癌的发生发展过程.     

Survivin在胆管癌组织中的表达及其与VEGF关系的相关研究

目的: 探讨凋亡抑制蛋白Survivin在胆管癌组织中的表达及其与血管内皮生长因子(vascular endothelial growth factor,VEGF)表达的关系.方法:应用免疫组织化学方法测定51例胆管癌组织和10例正常胆管组织的Survivin和VEGF的表达水平.结果:1)胆管癌组织中,Survivin阳性表达率为74.5%,正常胆管组织中的阳性表达率为0,两者相比差异有统计学意义(P＜0.01);Survivin在胆管癌中的表达与患者的性别和年龄、肿瘤的分化程度和部位、有无淋巴结转移无明显相关性(P＞0.05).2)VEGF在胆管癌组织中的阳性表达率为80.4%,在正常胆管组织中的阳性表达率为20%,两者相比差异有统计学意义(P＜0.01); VEGF在胆管癌中的表达与患者的性别和年龄、肿瘤的分化程度和部位无明显相关性(P＞0.05),而与有无淋巴结转移有相关性(P＜0.05),转移组高于无转移组.3)Survivin和VEGF在胆管癌组织中阳性表达的比率均为70.6%,二者的表达密切相关(P＜0.01).结论: Survivin和VEGF在胆管癌的发生、发展、侵袭和转移过程中发挥重要作用,二者在肿瘤血管生成过程中密切相关,为胆管癌的诊断和靶向基因治疗提供新的途径.     

凋亡抑制蛋白基因在先天性胆总管囊肿和胆管癌中的表达及意义

目的 探讨凋亡抑制蛋白基因(survivin)在先天性胆总管囊肿和胆管癌组织中的表达及意义.为进一步研究胆总管囊肿癌变的机制奠定理论基础.方法 采用免疫组化SP法检测Survivin在37例先天性胆总管囊肿组织(其中儿童20例,成人17例)和10例胆管癌组织中的表达,10例正常胆管作为对照.运用SPSS14.0统计学软件处理数据.结果 Survivin在37例先天性胆总管囊肿组织中,11例为阴性,15例弱阳性表达,9例阳性表达,2例强阳性表达.而在10例胆管癌组织中,Survivin有5例阳性和5例强阳性表达,未见阴性表达.正常胆管组织中仅有1例弱阳性表达,两两相比差异有统计学意义(P＜0.01).Survivin在成人胆总管囊肿中的表达强于儿童组,两者间差异有统计学意义(P＜0.01).结论 Survivin参与了胆总管囊肿癌变过程,可能与癌变的发生发展有关,在先天性胆总管囊肿癌变过程中可能起到促进作用.     

骨桥蛋白在胆管癌中的表达及其与抗药性的关系

目的:探讨肝胆管癌组织中骨桥蛋白的表达及其与抗药性的关系.方法:(1)采用实时定量PCR检测胆管癌及正常胆管组织骨桥蛋白mRNA的表达,并分析其表达与临床病理特征的关系;(2)应用靶向RNA干扰骨桥蛋白基因的技术构建骨桥蛋白低表达的细胞模型,并以CCK-8法检测5-FU对RBE胆管癌细胞增殖的抑制作用.结果:(1)胆管癌中骨桥蛋白的阳性表达率明显高于正常胆管组织(P＜0.01),骨桥蛋白mRNA表达水平与胆管癌的分化程度、肿瘤分期和淋巴结转移有关,上述指标的分组差异有统计学意义(P＜0.01),而与年龄、性别、肿瘤类型和病理类型等无明显关系,各组间比较差异无统计学意义(P＞0.05).(2)经沉默骨桥蛋白基因的RBE胆管癌细胞的骨桥蛋白mRNA表达显著降低(P＜0.01);且5-FU和顺铂抑制肿瘤细胞生长的IC50均显著降低(P＜0.01).结论:骨桥蛋白在胆管癌的发生、发展及产生抗药性方面起着一定的作用.     

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 comparison of the inhibitory effects of four volatile anaesthetics on the metabolism of chlorzoxazone, a substrate for CYP2E1, in rabbits

Background: Halothane inhibits in vitro and in vivo activity of cytochrome P-450 (CYP) 2E1. There are several fluorinated volatile anaesthetics besides halothane, and most of them are defluorinated by CYP2E1. It is unclear whether other fluorinated anaesthetics inhibit the in vivo activity of CYP2E1.
Methods: We compared the inhibitory effects of therapeutic concentrations of four inhalational anaesthetics, halothane, enflurane, isoflurane, and sevoflurane, on chlorzoxazone metabolism in rabbits receiving artificial ventilation.
Results: All four inhalational anaesthetics decreased arterial blood pressure and increased plasma chlorzoxazone concentration. However, no significant differences in the plasma chlorzoxazone concentration were found between the four anaesthetics. The estimated chlorzoxazone clearance increased after beginning inhalation with all four agents, but no significant difference in clearance was noted between agents.
Conclusions: At therapeutic concentrations, the in vivo inhibitory effect on chlorzoxazone metabolism was similar for all four inhalational anaesthetics examined, even though their chemical characteristics and extent of hepatic metabolism differ considerably.     

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.     

Is the recovery profile of mivacurium independent of the rate of decay of its plasma concentration in patients with normal plasma cholinesterase activity?

Background: The duration of action of muscle relaxants is poorly correlated to the rate of decay of their plasma concentration. The plasma concentration of mivacurium may rapidly decrease below its active concentration because of the extensive hydrolysis of mivacurium. By inflating a tourniquet on one upper limb for 3 min after the administration of atracurium, mivacurium or vecuronium, we studied the influence of the initial decline of their plasma concentration on their effect. Methods: In 50 patients anaesthetised with thiopental, isoflurane and fentanyl, the effect of bolus doses of 0.15 or 0.25 mg . kg^?1 mivacurium (MIV 15, MIV 25), 0.3 or 0.5 mg . kg^?1 atracurium (ATR 30, ATR 50) and 0.06 or 0.1 mg . kg^?1 vecuronium (VEC 06, VEC 10) were measured on both arms (evoked response of the adductor pollicis to train-of-four stimulation every 12 s), a tourniquet being applied on one arm just before and during 3 min after the muscle relaxant bolus. Results: Tourniquet inflation of 3 min almost abolished the neuromuscular effect of mivacurium. In the vecuronium groups and in the ATR 50 group, tourniquet inflation did not modify the maximum degree of depression of the twitch response. Also, the duration of action of vecuronium was unaffected by the tourniquet. In the ATR 30 group, times to return of the twitch response to 25% (duration 25%) and 75% (duration 75%) of control response were significantly shorter in the cuffed arm, 23 min vs 27 min, and 41 min vs 45 min, respectively. In the ATR 50 group, only duration 25% was significantly shorter in the cuffed arm (41 min vs 45 min). Conclusion: The results suggest that the rate of decline of the plasma concentration of mivacurium is so rapid, that a very low and almost clinically ineffective concentration is present as soon as 3 min after its administration. The results also indicate that the recovery from a mivacurium-induced neuromuscular blockade is not influenced by the rate of decay of its plasma concentration in patients with genotypically normal plasma cholinesterase.     

Membranes in Artificial Organs

Abstract: Membrane processes play a pivotal and enabling role in modern replacement therapy for acute and chronic organ failure and in the management of immunologic diseases. In fact, virtually all contemporary extracorporeal blood purification methods employ membrane devices, and the next generation of artificial organs and tissue engineering therapies are almost certain to be similarly grounded in membrane technology. In this short essay, we comment on the similarities and differences among synthetic membranes and their natural counterparts and also provide a critical overview of the demographics and technology of hemodialysis, hemofiltration, apheresis, oxygenation, and emerging membrane technologies and applications.     

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.     

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.     

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.     

Dilemma of Membrane Biocompatibility and Reuse

Abstract: Numerous articles have been published on the multiple use of dialyzers and on the effect of different reprocessing chemicals and techniques on the dialyzer biocompatibility and performance. The results often appear contradictory, especially those comparing standard biocompatibility parameters. Despite this confusion, a discerning review of the published works allows certain limited conclusions to be drawn. Reprocessing of used hemodialyzers changes the biocompatibility profile of a dialyzer as defined by the parameters complement activation. leukopenia, and cytokine release. The effect of reprocessing depends on the chemicals and reprocessing technique applied and also on the type of membrane polymer being subjected to the reprocessing procedure. Reports of pyrogenic reactions indicate that the flux of the membrane also influences how suitable it is for safe reuse. An increased risk of allergic and pyrogenic reactions appears to be associated with dialyzer reuse. Furthermore, there has been a lack of investigations into the immunologic effect of the layer of adsorbed and chemically altered proteins that remains on the inner surface of reprocessed dialyzers. We conclude that the clinical benefit of dialyzer reuse cannot be generally accepted from a biocompatibility point of view.