凋亡抑制因子XIAP在膀胱癌组织中的表达及其临床意义

目的检测抗凋亡（IAP）家族中XIAP在膀胱癌组织及癌旁正常组织的表达，探讨XIAP的表达在膀胱癌发生发展中的意义。方法采用免疫组织化学和实时荧光定量逆转录．聚合酶链反应（RT-QPCR）对50例膀胱癌患者中XIAP基因在癌组织和癌旁正常组织中的表达进行检测。结果免疫染色标本中，在空白对照、癌旁正常组织和膀胱癌组织中XIAP的阳性表达率分别为0、20％和60％。RT-QPCR显示XIAP在膀胱癌组织中的-△△CT值是癌旁组织的5．7163（4．3081-7．1245）倍，与分级、分期和有无转移没有相关性，但与是否复发有相关性。结论XIAP在癌旁正常组织中有少量表达，而在膀胱癌组织中的表达量高于癌旁组织，其表达量的多少可提肿瘤是否复发。     

葡萄糖调节蛋白94在人肝癌组织中的表达及其临床意义

目的 探讨葡萄糖调节蛋白94(Crp94)mRNA及蛋白在肝癌组织和癌旁组织中的表达情况及其与肝癌发生的关系.方法 肝癌患者手术切除标本10例,应用半定量逆转录-聚合酶链反应(RT-PCR)分别检测肝癌组织和癌旁组织细胞Grp94 mRNA基因的相对表达量,并通过免疫组织化学SP法检测10例肝癌组织和癌旁组织标本Crp94蛋白的表达情况.结果 (1)半定量RT-PCR显示:Grp94 mRNA在肝癌组织中均呈高表达,相对表达量为0.7490±0.7508,而在对应的癌旁组织中呈低表达或无表达,相对表达量为0.446 62±0.50000两组之间差异有统计学意义(P《0.05).(2)免疫组织化学及图像分析结果显示:在肝癌组织和癌旁组织中Grp94蛋白平均灰度扫描值分别为158.0800±20.5952、122.2800±24.4148两组之间差异有统计学意义(P《0.01).结论 Grp94在肝癌中的表达明显增高,可能在肝癌的发生、发展过程中发挥重要作用.     

EphrinA1在原发性肝细胞癌中的表达及临床意义

目的探讨原发性肝细胞癌中EphrinA1的mRNA和蛋白表达水平及其临床意义。方法应用逆转录聚合酶链反应（RT-PCR）及免疫组织化学方法检测EphrinA1的mRNA及蛋白在40例肝癌及相应的癌旁肝组织和10例正常肝组织中的表达水平,并分析其与临床病理学特点之间的关系。结果40例肝癌组织及相应的癌旁肝组织和10例正常肝组织中均有EphrinA1 mRNA的表达。RT-PCR分析显示EphrinA1mRNA在肝癌组织（0．5413±0．1527）中的表达显著高于癌旁肝组织（0．3895±0．0549,P〈0．05）和正常肝组织（0．3770±0．1055,P〈0．05）,而在癌旁肝组织（0．3895±0．0549）和正常肝组织（0．3770±0．1055）中的表达差异无统计学意义（P〉0．05）。从正常肝组织、癌旁肝组织到肝癌组织,EphrinA1蛋白的阳性表达率分别为20％（2／10）、35％（14／40）和62％（25／40）,呈递增趋势（x^2=14．762,P〈0．05）。EphrinA1蛋白的过表达与肝癌细胞的分化程度、门静脉癌栓的形成及淋巴结转移等临床病理因素有关（P〈0．05）。结论EphrinA1蛋白的过表达与肝癌细胞的分化程度、淋巴结转移和门静脉癌栓有密切联系,提示其可能在肝癌的恶性转化、侵袭和转移过程中发挥重要作用。     

人肝癌组织中核内转录基因Fos B和Jun D的表达及其意义

目的　探讨Fos和Jun的表达产物与人肝细胞癌发生、发展的关系。　方法　采用免疫组织化学S-P法和蛋白电泳技术对20例肝癌病人癌及癌旁组织中Fos和Jun核内癌蛋白的表达情况进行检测。结果　Westernblot结果示20例标本中有16例肝癌组织FosB、JunD蛋白含量高于癌旁组织。FosB光密度值在肝癌及癌旁组织中分别为2396±803和894±319,差异有显著意义（P＜0.01）;JunD光密度值在肝癌及癌旁组织中分别为1973±725和776±161,差异有显著意义（P＜0.05）。免疫组织化学结果显示FosB和JunD蛋白在肝癌组织细胞核中同步表达。　结论　在人原发性肝细胞癌组织中,核内转录基因FosB、JunD表达产物表达强度明显增高,这可能是肝细胞早期恶变的重要原因之一。     

凋亡抑制因子Livin在膀胱移形细胞癌中的表达

目的检测抗凋亡（IAP）家族中Livin基因在膀胱移形细胞癌（BTCC）组织及癌旁组织的表达，探讨Livin的表达在膀胱癌发生发展中的意义。方法采用免疫组织化学和实时荧光定量逆转录．荧光定量聚合酶链反应（RT-QPCR）方法对30例膀胱癌患者中Livin基因在癌组织和癌旁组织中的表达进行检测。结果免疫染色标本中，在癌旁组织和膀胱癌组织中Livin的阳性表达率分别为0．60％。Livin在膀胱癌组织中的-△△CT值是癌旁组织的8．0454（7．4264—8．6644）倍，与分级和分期没有相关性。结论Livin基因在癌旁组织中有少量表达，而在BTCC组织中的表达量远远高于癌旁组织。     

人垂体瘤转化基因在非小细胞肺癌组织中的表达

目的 从mRNA和蛋白质水平探讨人垂体瘤转化基因（hPTTG）表达和非小细胞肺癌（NSCLC）的关系，为肺癌基因诊断和基因治疗的研究提供理论依据。方法 应用逆转录-聚合酶链反应（RT-PCR）检测38例肺癌组织和癌旁正常肺组织中hPTTG mRNA的表达，再应用免疫组织化学方法和Western blot检测肺癌组织中hPTTG蛋白的表达，同时对实验对象进行随访资料生存分析，绘制生存率曲线，探讨与肺癌患者生存时间的关系。结果 RT-PCR显示肺癌组织中hPTTG mRNA表达明显高于癌旁正常肺组织（P〈0．05），不同类型和不同分化程度肺癌组织中hPTTG mRNA水平差异有统计学意义（P〈0．05），Ⅲ期肺癌组织中hPTTG mRNA表达明显高于Ⅰ期＋Ⅱ期（P〈0．05）；免疫组织化学显示肺癌组织中hPTTG蛋白表达明显高于癌旁正常肺组织（P〈0．05）；腺癌组织中hPTTG蛋白表达高于鳞癌组织；Western blot显示肺癌组织中hPTTG蛋白表达明显高于癌旁正常肺组织；生存分析方法显示hPTTG mRNA高表达者生存时间短于低／无表达者（P〈0．05，n=36）；用噻唑蓝（MTT）比色法观察生长曲线检测数据表明hPTTG的反义RNA表达载体转染组细胞的增殖明显比对照组细胞和空载体转染组细胞缓慢。结论 hPTTG表达增加与NSCLC发生、发展、浸润及转移有关，可作为判断NSCLC生物学行为的参考指标；hPTTG的反义RNA表达载体对SPC-A1肺癌细胞株的生长具有一定的抑制作用。     

凋亡抑制基因Livin在食管癌中的表达及其与P53、Bcl-2的关系

目的探讨凋亡抑制基因Livin在食管癌组织中的表达及其与P53和Bcl-2表达的关系。方法采用逆转录-聚合酶链反应（RT—PCR）结合银染技术检测36例食管癌组织和18例癌旁正常组织中Livin mRNA的表达，采用免疫组织化学链霉菌抗生物素蛋白-过氧化酶（S—P）法检测食管癌组织中Livin、P53和Bcl-2蛋白的表达。结果RT—PCR检测结果显示：Livin mRNA在食管癌组织中的阳性表达较癌旁正常组织明显增强，两种异构体基本同时表达。免疫组织化学检测结果显示：Livin蛋白在食管癌组织中的阳性表达率明显高于癌旁正常组织（P〈0.01）。癌组织侵及食管外膜的Livin阳性表达率高于癌组织侵及食管肌层者（P〈0.05）；有淋巴结转移者的Livin阳性表达率高于无淋巴结转移者（P〈0.05）。Livin蛋白表达与P53蛋白表达无关（x^2=1．00，P=0．505），与Bcl-2蛋白表达有关（x^2=10．60，P=0．003）。结论Livin在食管癌组织中异常表达，有望成为食管癌诊断和基因治疗的新靶点。凋亡相关基因Bcl-2与Livin基因的异常表达可能在食管癌癌变中起协调作用。     

促凋亡因子Omi/HtrA2对凋亡抑制蛋白表达及A549细胞凋亡的影响

目的观察Omi／HtrA2对凋亡抑制蛋白（XIAP）表达的影响和对肺癌A549细胞凋亡的影响。方法将构建的携带绿色荧光蛋白（GFP）和Omi／HtrA2基因的表达载体（PEGFP-N1-Omi）转染至人肺腺癌A549细胞中，用逆转录-聚合酶链反应（RT-PCR）和Western blot检测Omi／HtrA2对XIAP转录和表达的影响，流式细胞仪检测Oml／HtrA2协同顺铂作用于A549细胞后该细胞的凋亡变化。结果Omi／HtrA2基因转染组XIAP的mRNA表达水平下降37％（P〈0．05），XIAP的蛋白表达水平下降69％（P〈0．05），顺铂诱导后，基因转染组细胞凋亡率为（36．00±1．95）％，显著高于对照组的细胞凋亡率（17．00±1．12）％（P〈0．05）。结论Omi／HtrA2通过抑制XIAP的抗凋亡作用可协同顺铂促进A549细胞凋亡。     

Twist基因在肝细胞肝癌中的表达及意义

目的探讨转录因子Twist在人肝细胞肝癌、癌旁和肝硬化组织中的表达及其意义。方法利用免疫组化方法检测26例肝细胞肝癌标本（癌和癌旁组织）和10例肝硬化标本中Twist的表达情况;利用逆转录聚合酶链反应（RT—PCR）和Western蛋白印迹方法检测其中10例标本中的Twist mRNA和其蛋白的表达情况,分析其临床病理意义。结果免疫组化显示：Twist蛋白在肝癌、癌旁和肝硬化组织中的阳性率分别为84．6％、19．2％和20．0％,Twist在肝癌组织中的表达明显强于癌旁或肝硬化组织（P〈0．05）;大多数癌旁和肝硬化组织无Twist表达,癌旁与肝硬化组织相比阳性率无差别（P〈0．05）。与癌旁组织相比,Twist mRNA和其蛋白在肝癌组织中表达明显上调（P〈0．05）。结论Twist在肝细胞肝癌组织中的过高表达与肝癌的发生发展相关。     

肝癌及癌旁组织中β-葡萄糖醛酸酶蛋白及mRNA的表达及意义

目的探讨肝细胞癌及癌旁组织中β-葡萄糖醛酸酶（β-G）蛋白及β-G mRNA的表达和临床意义。方法采用逆转录-聚合酶链反应（RT-PCR）方法和免疫组织化学方法，对25例原发性肝细胞癌及其癌旁组织和10例正常肝脏组织进行检测，并分析其与临床病理特点之间的关系。结果半定量分析显示β-G mRNA的表达在肝脏癌组织（3．71±0．32）与癌旁肝组织（1.83±0.22）以及正常肝脏组织（1．71±0．32）间差异比较，均有统计学意义（均为P〈0．01），而癌旁肝组织和正常肝组织间比较，差异无统计学意义（P〉0．05）；从正常肝组织、癌旁组织到肝癌组织β-G蛋白的阳性细胞表达率分别为（48．2％±5．5％）、（56．8％±6．2％）和（83．8％±9．1％），呈递增趋势（χ^2=42．33，P〈0．01）；β-G蛋白的表达与肝癌病人的门脉癌栓以及淋巴结转移等临床因素有关（P〈0．01），而与肿瘤大小、AFP水平无关。结论β-G与肝细胞的癌变过程密切相关，可能在肝癌的侵袭和转移过程中也起一定的作用。     

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 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.     

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.     

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.