人脑胶质瘤中全长新基因PPIL3的获得及染色体定位和表达谱分析

目的运用基因芯片技术获取正常成人脑组织与人脑胶质瘤中差异表达的基因，并对其中一条基因进行研究。方法抽提正常成人脑组织与人脑胶质瘤组织中的mRNA来制备探针，经杂交、洗涤后，通过计算机观察两者表达谱的差异情况，对6811705克隆子进行Northern blot，逆转录-聚合酶链反应（RT-PCR），染色体定位和生物信息学分析。结果通过4次基因芯片筛选，获得15条与胶质瘤相关的新基因，经Northern blot证实6811705基因在人正常脑组织中低表达，而在人脑胶质瘤中高表达。PPIL3基因位于染色体2q33区段的STS marker stSG2762和SHGC-3074之间。BLASTn和BLASTx分析显示，它们编码蛋白与线虫Cyp-10蛋白同源性分别为52％和72％。cDNA序列分析发现这两上克隆是同一个基因[命名为eyelophilin-like gene（PPIL3）]的两个不同的剪切体（PPIL3a和PPIL3b）。结论基因芯片筛选正常脑组织与人脑胶质瘤差异表达的基因具有样品用量少，高质量，高速度，高敏感等特性。6811705基因可能是与人脑胶质瘤形成有关的一条全长新基因。     

全长新基因PPIL3的获得及在人脑胶质瘤中的表达

目的 运用基因芯片技术获取正常成人脑组织与人脑胶质瘤中差异表达的基因,并对其中1条基因进行初步研究.方法 抽提正常成人脑组织与人脑胶质瘤组织中的mRNA来制备探针,经杂交、洗涤后,通过计算机观察两者表达谱的差异情况,对681F05克隆子进行Northern blot,原位杂交和生物信息学分析.结果 通过4次基因芯片筛选,获得15条与胶质瘤相关的新基因,经northern blot证实681F05基因在人正常脑组织中低表达,而在人脑胶质瘤中高表达.原位杂交得到相同的结果.BLASTn和BLASTx分析显示,它们编码蛋白与线虫Cyp-10蛋白同源性分别为52%和72%.cDNA序列分析发现这两上克隆是同一个基因(命名为cyclophilin-like gene (PPll3))的两个不同的剪切体(PPIL3a和PPll3b).结论 基因芯片筛选正常脑组织与人脑胶质瘤差异表达的基因具有样品用量少,高质量,高速度,高敏感等特性.681F05基因可能是与人脑胶质瘤形成有关的一条全长新基因.     

人脑胶质瘤差异表达的基因韵获得及一条全长新基因的克隆

目的 运用基因芯片技术获取正常成人脑组织与人脑胶质瘤中差异表达的基因，并对其中一条基因进行了初步的研究。方法 抽提正常成人脑组织与人脑胶质瘤组织中的mRNA来制备探针，经杂交、洗涤后，通过计算机观察两者表达谱的差异情况，对681F05克隆子进行了Nozxhem blot，5’RACE和生物信息学分析。结果通过4次基因芯片筛选，获得15条与胶质瘤相关的新基因，经Northem blot证实681F05基因在人正常脑组织中低表达，而在人脑胶质瘤中高表达。BLASTn和BLASTx分析显示，它们编码蛋白与线虫Cyp-10蛋白同源性分别为52％和72％。cDNA序列分析发现这两上克隆是同一个基因[命名为cyclophilin-like gene(PPIL-3)]的两个不同的剪切体(PPIL3a和PPIL3b)。结论 基因芯片筛选正常脑组织与人脑胶质瘤差异表达的基因具有样品用量少，高质量，高速度。高敏感等特性。681F05基因可能是与人脑胶质瘤形成有关的一条全长新基因。     

人脑胶质瘤差异表达的基因的获得及一条全长新基因的克隆

目的运用基因芯片技术获取正常成人脑组织与人脑胶质瘤中差异表达的基因,并对其中一条基因进行了初步的研究。方法抽提正常成人脑组织与人脑胶质瘤组织中的mRNA来制备探针,经杂交、洗涤后,通过计算机观察两者表达谱的差异情况,对681F05克隆子进行了Northernblot,5’RACE和生物信息学分析。结果通过4次基因芯片筛选,获得15条与胶质瘤相关的新基因,经Northernblot证实681F05基因在人正常脑组织中低表达,而在人脑胶质瘤中高表达。BLASTn和BLASTx分析显示,它们编码蛋白与线虫Cyp10蛋白同源性分别为52%和72%。cDNA序列分析发现这两上克隆是同一个基因[命名为cyclophilinlikegene(PPIL3)]的两个不同的剪切体(PPIL3a和PPIL3b)。结论基因芯片筛选正常脑组织与人脑胶质瘤差异表达的基因具有样品用量少,高质量,高速度,高敏感等特性。681F05基因可能是与人脑胶质瘤形成有关的一条全长新基因。     

人脑胶质瘤中全长新基因PPIL3的生物信息学研究

目的 应用基因芯片技术获取正常成人脑组织与人脑胶质瘤中差异表达的基因,并对其中1条基因进行初步研究.方法 抽提正常成人脑组织与人腩胶质瘤组织中的mRNA制备探针,经杂交、洗涤后,通过计算机观察两者表达谱的差异,对681 F05克隆子进行Northern blot和生物信息学分析.结果 通过4次基因芯片筛选,获得15条与胶质瘤相关的新基因,经Northern blot证实681F05基因在人正常脑组织中低表达,而在人脑胶质瘤中高表达.BLASTn和BLASTx分析显示,它们编码蛋白与线虫Cyp-10蛋白同源性分别为52%和72%.cDNA序列分析发现此两个克隆是同一个基因[命名为cyclophilin-like gene(PPIL3)]的两个不同的剪切体(PPIL3a和PPIL3b).结论 基因芯片筛选正常脑组织与人脑胶质瘤差异表达的基因具有样品用量少、高质量、高速度、高敏感等特性.681F05基因可能是与人脑胶质瘤形成有关的一条全长新基因.

Abstract：

Objective To obtain differentially expressed genes related to human glioma using cDNA microarray and study the characterization of one novel full-length gene. Methods Total RNA was extracted from human glioma tissues and normal brain tissues, and mRNA was used to make probes. After hybridization and washing procedure, the results of hybridization were scanned using computer system. One gene named 681F05 clone was subsequently analyzed by northern blotting and bioinformatic analysis. Results We obtained 15 differentially expressed genes to human glioma through 4 times hybridizations and scanning. Northern blotting analysis confirmed 681F05 clone was down-regulated in human brain tissue and up-regulated in human glioma tissues. The analysis of BLASTn and BLASTx showed that clone 681F05 isolated was two cDNA clones encoding two novel proteins which showed 52% and 72% identity to the cyclophilin isoform 10 of C. Elgans, respectively. Sequence analysis revealed these two cDNA clones were two different splicing variants of a novel cycophilin-like gene ( PPIL3a and PPIL3b). Conclusion cDNA microarray technology can be successfully applied to identify differentially expressed genes. The novel fulllength gene of human PPIL3 may be correlated with tumorigenesis of human glioma.     

脑胶质瘤相关新基因PKIβ的表达与蛋白质活性研究

新基因,共编码78个氨基酸,其理论相对分子质量为8468 Da,等电点为4.69,与鼠PKI 69%同源,命名为人PIG.并在大肠杆菌中得到了PKI较高的表达蛋白,经纯化,在SDS-PAGE胶上获得1条清晰的条带.重组的PKIβ-78蛋白和PKIβ71蛋白均对PKA C亚基有明显抑制作用.结论 基因芯片筛选正常脑组织与人脑胶质瘤差异表达的基因具有样品用量少,高质量,高速度,高敏感等特性.人PKI可能是与人脑胶质瘤形成有关的一条全长新基因.     

应用高密度寡核苷酸(Oligo)基因芯片筛选胶质瘤相关基因

目的应用高密度寡核苷酸（Oligo）基因芯片技术筛选胶质瘤相关基因。方法抽提5例胶质瘤和10例正常脑组织的总RNA并逆转录为cDNA，其中Cy5、Cy3的dNTP分别掺人胶质瘤和正常脑组织的cDNA，混合后杂交含22000个人类基因人类高密度Oligo基因芯片，经过洗片和扫描，获得荧光信号图像并用计算机分析，随机抽取3种差异表达基因用PCR验证它们胶质瘤和正常脑组织中的差异表达。结果从22000条基因中筛选出差异表达基因242条，其中123条表达上调，119条表达下调，包括细胞周期蛋白、细胞凋亡、细胞骨架和运动蛋白等相关基因。结论基因芯片技术的胶质瘤基因表达谱分析能够高通量筛选胶质瘤相关基因，并高效对基因功能进行研究，有助于认识肿瘤发病机制。     

人脑胶质瘤中连接蛋白基因表达及其临床意义

目的探讨人脑胶质瘤连接蛋白(Cx)43、32基因表达及其临床意义。方法对8例正常脑组织和50例人脑胶质瘤采用Northern印迹和免疫组织化学法检测。结果人脑胶质瘤Cx43mRNA及其蛋白表达阳性率分别为54％、52％，Cx32仅在1例低恶度少枝胶质细胞瘤及1例混合性星形一少枝胶质细胞瘤中表达。Cx43mRNA及其蛋白表达水平随肿瘤恶性程度升高而下降。结论Cx基因在人脑胶质瘤中表达可下降或缺失，与肿瘤的病理类型及恶性程度呈负相关，可作为某些肿瘤的病理诊断参考指标之一。     

基因芯片检测人脑胶质瘤恶性进展的分子变化

目的　探讨胶质瘤恶性进展过程中的分子变化 ,为进一步研究胶质瘤的分子亚型作准备。方法　收集同一患者低级别向高级别发展过程中 3次手术标本及远离病灶的正常脑组织 ,用基因芯片技术检测、分子生物信息学分析、优筛表达差异有非常显著性、有进一步研究价值的基因 ;再用半定量逆转录 聚合酶链反链 (RT PCR)验证其在不同患者、不同级别手术标本中的表达。结果　在基因芯片中 ,4份标本两两对比后获得差异表达 2倍和 3倍以上的基因分别有 2 0 5 3条和489条 ,他们分布在 15类功能不同的基因组中 ,有些是不同级别特有的表达差异基因。筛选到与胶质瘤恶性进展相关的 4条基因 (X5 43 0 4、NM 19896、AI2 64 2 16、NM 15 962 ) ,在 2 2例临床标本中 ,经半定量RT PCR检测表明 ,其表达量随恶性程度增高而增加或减少。结论　胶质瘤基因芯片和RT PCR结合测得的与恶性程度相关的基因 ,可为进一步研究胶质瘤的分子病理亚型提供实验依据。     

苯乙酸对胶质瘤U-251细胞中RNA编辑酶表达的影响

目的观察正常人脑胶质细胞和胶质瘤U-251细胞中RNA编辑酶RED1,RED2 mR- NA水平的表达,并观察诱导分化剂苯乙酸对U-251细胞中RED1 mRNA表达的影响。方法对原代培养的正常人脑胶质细胞和胶质瘤U-251细胞,应用RED1,RED2全长序列合成引物及合成的特异引物,分别通过逆转录-聚合酶链反应(RT-PCR)检测RED1,RED2 mRNA水平的表达。用RT- PCR及图像分析法,检测胶质瘤细胞U-251在不同浓度的苯乙酸处理前后,RED1 mRNA表达水平的变化。RED1基因表达水平用基因/β-肌动蛋白(β-actin)灰度比值表示。结果RED1在正常人脑胶质细胞中表达极弱,在高恶性度的胶质瘤U-251细胞中明显表达。诱导分化剂苯乙酸作用后, U-251细胞中RED1表达水平降低。RED2在正常人脑胶质细胞及苯乙酸处理前后的胶质瘤细胞中,均未见表达。结论RED1 mRNA水平高表达,可能与高恶性度胶质瘤的发生有关。诱导分化剂苯乙酸可能通过降低RED1mRNA水平的表达,作用于胶质瘤细胞的RNA编辑过程。     

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.