凋亡抑制蛋白Survivin在骨肉瘤中的表达及其与PTEN、cyclinD1蛋白表达的关系

[目的]探讨Survivin基因的表达在骨肉瘤发生、发展中的作用及其与PTEN、cyclinD1蛋白表达的相互关系。[方法]应用免疫组化（SP）法检测40例骨肉瘤组织、20例骨软骨瘤组织中Survivin、PTEN、cyclinD1蛋白的表达情况以及Survivin与PTEN、cyclinD1蛋白两者之间的相关性。[结果]Survivin在骨肉瘤中的阳性率为62．5％（25／40），明显高于对照组骨软骨瘤10．0％（2／20）的阳性率（P〈0．01）。PTEN蛋白在骨肉瘤中的阳性表达率为52．5％（21／40），而在骨软骨瘤中为95．0％（19／20），两者相比有显著差异（P〈0．01）。cyclinD1蛋白在骨肉瘤中的阳性率为80．0％（32／40），高于在骨软骨瘤中的45．0％（9／20），有统计学意义（P〈0．05）。Survivin和PTEN两者表达强度之间呈显著负相关（P〈0．01），而Survivin与cyclinD1表达呈正相关（P〈0．01）。[结论]Survivin在骨肉瘤组织中过表达与骨肉瘤的发生有关；PTEN在骨肉瘤组织中失表达，它可能下调Survivin表达，对骨肉瘤起抑制作用：Survivin和cyclinD1在骨肉瘤的发生发展中可能起协同作用。     

转移抑制基因maspin在骨肉瘤中的表达及其意义

目的探讨肿瘤转移抑制基因maspin在骨肉瘤中的表达及意义。方法收集自1996年至2004年在中国医科大学附属第一医院骨科诊治的骨肉瘤患者的完整资料及手术标本，以及术后保留的瘤旁正常骨组织，共62例。骨肉瘤标本依据Enneking分期分组，分别予1％多聚甲醛固定和深低温冷藏及脱钙。采用逆转录一聚合酶链反应（RT-PCR）和免疫组织化学方法（SP法）检测maspin基因在骨肉瘤和正常骨组织中的表达，探讨其表达与骨肉瘤的临床特性之间的关系。结果RT-PCR：maspin mRNA在正常骨组织中的表达量明显高于在骨肉瘤中的表达量，组间差异有统计学意义（P〈0．05）。免疫组织化学：maspin在骨肉瘤和正常骨组织中的阳性率分别为76．1％和59．5％，组间差异有统计学意义（P〈0．05）。maspin基因的表达与骨肉瘤患者的临床分期呈负相关（P〈0．05）。结论转移抑制基因maspin的表达下调在骨肉瘤发展中起着一定作用，maspin的表达变化可能对骨肉瘤的转移具有抑制作用。     

细胞黏附分子在骨肉瘤中的表达及其临床意义

[目的]检测CD44V6、β-catenin在骨肉瘤组织中的表达，分析其表达与人骨肉瘤细胞浸润及转移能力的相关性。[方法]应用免疫组织化学方法检测35例骨肉瘤、15例骨软骨瘤、15例正常骨组织中CD44V6、β-catenin的表达。[结果]骨肉瘤组织中CD44V6、β-catenin的阳性表达显著高于骨软骨瘤及正常骨组织（P〈0．01）；两者的表达分别与骨肉瘤临床分期、肿瘤转移及2年内死亡有关（P〈0．05），而与患者性别、年龄、肿瘤大小、病理分型、术前化疗无关（P〉0．05）；两者的阳性表达呈正相关（P〈0．01）。[结论]CD44V6、β-catenin在骨肉瘤的阳性表达与骨肉瘤的发生、发展和转移密切相关，可能在骨肉瘤细胞侵袭、转移的过程中起协同作用；联合检测CD44V6、β-catenin可以作为骨肉瘤的诊断和预测转移的指标。     

骨肉瘤中血管生成素-2与受体Tie-2 mRNA表达及其意义

目的 探讨血管生成素-2（Ang-2）及其受体Tie-2在骨肉瘤血管生成中作用及其对骨肉瘤恶性演进的影响。方法 采用原位杂交和免疫组织化学技术，检测46例骨肉瘤、15例骨软骨瘤、5例正常骨组织中Ang-2、Tie-2mRNA表达及骨肉瘤中VEGF、CD34蛋白表达，计数微血管密度（MVD）；并与骨肉瘤主要病理学参数进行比较。结果 Ang-2、Tie-2mRNA在正常骨组织无表达，少量表达于骨软骨瘤及骨肉瘤瘤旁组织，明显表达于骨肉瘤中（69．6％、73．9％）；Ang-2、Tie-2表达水平与骨肉瘤组织学分级分型无关，转移组显著高于无转移组（P〈0．01）；高MVD组高于低MVD组（P〈0．01），Ang-2与VEGF表达相关，Ang-2与Tie-2表达正相关（r=0．445，P〈0．01）。结论 Ang-2及其受体Tie-2参与了骨肉瘤血管生成调控，与骨肉瘤浸润性生长和转移密切相关。     

MHCⅠ类链相关蛋白A和基质金属蛋白酶-9在骨肉瘤组织学和血清学中的关系

目的 探讨MHCI类链相关蛋白A（MHC class I chain-related A,MICA）和基质金属蛋白酶-9（matrix metalloproteinase-9,MMP-9）在骨肉瘤组织学和血清学中的关系,为研究骨肉瘤组织中MICA蛋白脱落的分子机制提供组织学基础。方法应用免疫组织化学方法检测16例骨肉瘤中MICA和MMP-9蛋白的表达情况,同时采用酶联免疫吸附实验（enzyme-linkedimmunosorbentassay,ELISA）检测16例骨肉瘤患者血清中MICA和MMP-9的含量。结果 16例骨肉瘤组织中MICA和MMP-9蛋白的表达率分别为42．5％（7／16）和56．3％（9／16）,两者的表达无相关性（r=0．454,P〉0．05）;骨肉瘤患者血清中可溶性MICA（solubleMICA,sMICA）浓度高于健康人（P〈0．05）,与血清中MMP-9的浓度无相关性（r=-0．429,P〉0．05）,但与组织中MICA蛋白表达呈负相关（r=-0．502,P〈0．05）。结论骨肉瘤患者血清中sMICA浓度升高,而血清中MMP-9含量对MICA浓度无明显影响;骨肉瘤患者可能存在NKG2D-MICA介导的免疫监视功能障碍,MICA蛋白的脱落可能是骨肉瘤逃避免疫监视的一个新机制。     

ras-MAPK信号通路在骨肉瘤中的表达及其意义

目的探讨骨肉瘤中ras—MAPK信号传导通路的相互关系，观察它们在骨肉瘤发生、发展中的作用。方法选取我科自2000年至2006年40例骨肉瘤手术标本，标本在切下后转入深低温保存。分别应用免疫组织化学和Westernblot检测在骨肉瘤和正常骨组织中的raS基因产物p21和MAPK信号通路标志蛋白p38的表达，并分析在ras p21不同表达的骨肉瘤组织中的p38的表达，探讨两者之间的相关性。结果免疫组织化学和蛋白检测结果显示，所有40例标本中ras p21、p38在骨肉瘤组织中的蛋白表达量与正常骨组织之间差异有统计学意义，骨肉瘤中的表达均明显高于正常骨（P〈0、05）；rasp21与p38的表达呈现明显的正相关：在ras p21含量较高的骨肉瘤组织中p38的含量较高，在ras p21含量较低的骨肉瘤组织中p38的含量较低（P〈0．05）。结论骨肉瘤中存在着ras—MAPK信号传导通路的表达，该传导通路的激活在骨肉瘤的发生、发展中起到一定作用。     

表皮生长因子受体和蛋白激酶B在胃癌组织中的表达及临床意义

目的探讨胃癌组织表皮生长因子受体（EGFR）和蛋白激酶B（AKT）在胃癌组织中的表达与胃癌临床病理特征及预后的关系。方法采用免疫组织化学方法检测84例胃癌组织和15例非癌胃组织中EGFR和AKT的表达情况,并分析其表达与胃癌患者临床病理特征及预后的关系。结果EGFR和AKT在胃癌组织的阳性表达率分别为55．9％（47／84）和64．3％（54／84）,明显高于非癌胃组织的20．0％（3／15）和6．7％（1／15）,差异均有统计学意义（P〈0．05,P〈0．01）。EGFR表达与胃癌分期有关,AKT表达则与胃癌淋巴结转移有关（均P〈0．01）。EGFR阴性表达和阳性表达患者5年总体生存率分别为49％和22％（P〈0．05）;AKT阴性表达和阳性表达患者5年总体生存率分别为45．7和30．2％（P〈0．05）。结论检测胃癌组织中的EGFR和AKT的表达,有助于判断胃癌恶性程度及评估预后。     

人骨肉瘤中S100A6蛋白的表达及其意义

目的观察人骨肉瘤中S100A6蛋白的表达,探讨其与肿瘤转移的关系以及对预后的影响。方法采用免疫组织化学技术对30例骨软骨瘤和60例骨肉瘤石蜡包埋标本进行检测,对9例骨软骨瘤和7例骨肉瘤泳冻标本进行Western-blot检测,分析其表达与转移和预后的关系。结果与骨软骨瘤（10％）相比,S100A6蛋白显著表达于人骨肉瘤（85％）中（P〈0．01）,其表达与性别、肿瘤的分期、分型无关（P〉0.05）,而与肿瘤的转移（P〈0．05）和术后生存时间（P〈0．01）密切相关。结论S100A6蛋白选择表达于骨肉瘤中,可作为肿瘤的早期诊断标志之一,也是判断骨肉瘤转移和预后的重要指标。     

PTEN和p27及cyclin D1蛋白在大肠癌组织中的表达及相关性研究

目的：研究大肠癌组织中PTEN、p27和cyclinD1蛋白的表达及三者间的关系。方法：应用免疫组铡匕学方法检测58例大肠癌组织、58例癌旁组织和14例正常大肠黏膜中PTEN、p27和cyclinD1蛋白的表达情况。结果：大肠癌组织中PTEN和p27蛋白的阳性表达率分别为65．52％（38／58）和53．45％（31／58），较癌旁组织和正常组织明显刚氏（P〈0．05）；cyclinD1蛋白的阳性表达率为60．34％（35／58）,较癌旁组织及正常组织显著升高（P〈0．05）。三者在大肠癌组织中的表达存在相关性，且均与肿瘤的分化程度、有无淋巴结转移和Dukes分期有关（P〈0．05）。结论：PTEN、p27和cyclinD1基因的异常改变参与大，且三者之间存在相关性。联合检测三者表达水平可作为评价大肠癌病理生物学行为的客观指标之一。     

DOG-1蛋白在胃肠间质瘤中的表达及其意义

目的探讨DOG．1在胃肠间质瘤（GIST）中的表达及其与GIST临床病理特征的关系。方法应用组织芯片技术及免疫组织化学方法检测80例GIST及40例正常胃肠道组织中DOG-1及CD117的表达情况。结果80例GIST组织中有76例（95．0％）表达DOG．1,67例（83．8％）表达CD117,两者比较差异有统计学意义（P〈0．05）;而40例正常胃肠道组织中,均未见CD117及DOG．1阳性表达。梭形细胞型GIST中,DOG．1与CD117阳性表达率均为96．0％（49／51）：上皮样细胞型GIST两者表达率分别为94．1％（16／17）和52．9％（9／17）;混合细胞型GIST分别为91．7％（11／12）和75．0％（9／12）。DOG．I的阳性表达与GIST患者的年龄、性别、肿瘤的发生部位及病理学分级均无关（均P〉0．05）。结论DOG．1在GIST组织中高表达。特别是在以上皮样细胞为主型中存在高表达,在正常胃肠道组织中不表达,有望作为诊断GIST的一种新型的标志物。     

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.     

Systemic analgesia adapted to the children's condition

A concept of balanced analgesia using nonsteroidal anti-inflammatory drugs (NSAIDs), paracetamol (acetaminophen), opioids, and corticosteroids can also be used in patients with pre-existing illnesses. NSAIDs are the most effective treatment for acute pain of moderate intensity in children; however, these drugs should be avoided in patients at increased risk for serious side effects, e.g. patients with renal impairment, bleeding tendency, or extreme prematurity. NSAIDs can be given with minimal risks to the younger child with mild to moderate asthma, and, in these patients, the use of steroids can be encouraged; in addition to their antiemetic and analgesic action, a beneficial effect on asthma symptoms can be expected. In the non-intubated child with cerebral trauma, exaggerated sedation caused by opioids and increased bleeding tendency caused by NSAIDs must be avoided. In neonates and small infants, the oral administration of sucrose or glucose is helpful to minimize pain reaction during short uncomfortable interventions.