脂质体介导的端粒酶反义寡核苷酸在人膀胱癌EJ细胞中的转染率及稳定性

目的探讨端粒酶反义寡核苷酸（asODN）在脂质体介导下在入膀胱癌EJ细胞中的转染率、分布特点及稳定性。方法合成针对端粒酶RNA模板区的asODN，并行全硫代修饰或不修饰，5-FITC标记，脂质体介导转染人膀胱癌EJ细胞后分别于15、30min、1h以及以后每小时，在电镜下观察asODN在细胞中的表达，分布特点及稳定性。结果在脂质体介导下，未修饰型asODN转染细胞后15min开始表达，1～3h稳定表达于细胞中，表达率15％～20％，4h减退；修饰型asODN 15min开始表达，8h有56％～80％的高表达，12h开始消散。结论脂质体是一种有前途的膀胱癌非病毒基因治疗载体。     

端粒酶反义RNA基因转染抑制裸鼠人膀胱癌移植瘤生长的研究

目的　观察端粒酶反义RNA基因转染对裸鼠膀胱癌移植瘤细胞生长的影响。方法将脂质体介导的端粒酶反义RNA真核表达载体 (pBBS hTR)、空载体 ( pBBS 2 12 )及生理盐水注入移植瘤体内 (每天 3 0 0 μl,共 7d) ,应用端粒酶活性聚合酶链反应 酶联免疫吸附试验 (PCR ELISA)、苏木素 伊红 (HE)染色、透射电镜等连续观察 6周移植瘤组织细胞端粒酶活性和生长变化。结果　注射转染pBBS hTR组和空载体组瘤体积抑制率分别为 48.7%和 2 .8% (P <0 .0 5 )。转染pBBS hTR组端粒酶活性降低 ,细胞生长受抑制。电镜观察见典型凋亡特征。 结论　端粒酶反义RNA基因瘤体内注射转染有效地抑制裸鼠人膀胱癌细胞生长 ,具有潜在临床应用价值。     

端粒酶反义RNA转染促进膀胱癌T24细胞凋亡的研究

目的　探讨端粒酶反义RNA对膀胱癌T2 4细胞恶性表型的抑制及促进其凋亡的作用。　方法　采用脂质体转染法将转录出端粒酶反义RNA质粒导入膀胱癌T2 4细胞。应用PCR ELISA法测定转染后T2 4细胞的端粒酶活性 ;光镜、电镜、MTT及流式细胞术 (FCM )等方法观察端粒酶反义RNA对T2 4细胞生长及凋亡的影响。　结果　端粒酶反义RNA能显著抑制T2 4细胞的端粒酶活性 ,转染T2 4细胞后使其生长受到抑制。形态学观察 ,转染后T2 4细胞出现典型的凋亡现象。FCM检测发现G1期前出现凋亡峰。　结论　转染端粒酶反义RNA能抑制膀胱癌T2 4细胞的恶性表型 ,促进其凋亡     

抑制端粒酶活性诱导膀胱癌细胞凋亡的研究

目的 探讨抑制端粒酶粒酶活性各市县导膀胱癌细胞凋亡的可能性。方法 采用具有5′-d(TTAGGG)-3′序列的硫代磷酸反义寡核苷酸（PS-ODN）与膀胱癌EJ细胞共培养,观察细胞内端粒酶活性变化。细胞生长状态及细胞凋亡形态学变化。结果 经PS-ODN处理的膀胱癌细胞内端粒酶活性下降或消失,细胞生长状态受到明显抑制,并出现细胞凋亡特有的形态变化。结论 具有5′-d(TTAGGG)-3′序列的PS-ODN能够抑制膀胱癌细胞内的端粒酶活性,抑制膀胱癌细胞生长,诱导膀胱癌细胞凋亡。     

端粒酶反义寡核苷酸对乳腺癌端粒酶活性及细胞生长的影响

目的　探讨端粒酶反义寡核苷酸对人乳腺癌细胞端粒酶活性的影响及抑制端粒酶活性以控制乳腺癌细胞生长的可能性。方法　采用 10 μmol/L与端粒酶催化亚基 (hTRT )mRNA互补的反义寡核苷酸处理乳腺癌细胞 ;于处理后 2 4、72h用端粒重复序列扩增及酶联免疫吸附方法检测细胞端粒酶活性 ;于处理后 72h以流式细胞术测定细胞周期 ;于处理后 12 0h采用原位末端标记法检测细胞凋亡。结果　经hTRT反义寡核苷酸作用后 ,细胞端粒酶活性明显受到抑制 ,至作用后 72h ,端粒酶活性较对照组降低 65 .6% (P <0 .0 5 ) ;细胞生长明显受到抑制 ;细胞周期发生显著变化 :G0 /G1期细胞数增多 ,S期及G2 /M期细胞数则减少 ;细胞增殖指数由 0 .46降低至 0 .2 5 ;并可观察到凋亡细胞的出现 ,凋亡发生率为 12 .5 % ;而无义组及空白对照组以上各指标均无明显变化 (P >0 .0 5 )。结论　端粒酶反义寡核苷酸可明显抑制乳腺癌细胞端粒酶活性 ,抑制细胞生长和增殖 ,诱导细胞凋亡 ;表明应用反义技术抑制端粒酶活性治疗乳腺癌具有广阔的前景。     

超微载体介导反义端粒酶RNA抑制胶质瘤细胞生长的研究

目的　研究用体内可降解的聚乳酸 (PLA)和o 梭甲基壳聚糖 (CMC)制成的超微载体介导端粒酶RNA(hTR)反义寡核苷酸在体外对TJ90 5人脑胶质瘤细胞的作用。方法　用PLA和CMC制成超微载体 ,并用其介导hTR反义寡核苷酸在体外转染TJ90 5细胞 ,通过噻唑兰比色法(MTT)、改良端粒重复序列扩增法 (TRAP)、逆转录 聚合酶链反应 (RT PCR)对细胞转染情况作检测。结果　超微载体在体外能有效转染hTR反义寡核苷酸 ,48h后细胞存活率为 46.84% ,hTR、端粒酶催化亚基mRNA和端粒酶的活性水平分别为 0 .3 16、0 .0 2 4、5 1.40 0 ,明显受到抑制。结论　hTR可作为胶质瘤基因治疗靶点 ,超微载体能有效转染基因药物 ,可替代病毒载体。     

A型精原细胞体外分离纯化及其端粒酶活性抑制后的形态学变化

目的 :探讨分离纯化、体外培养的A型精原细胞在端粒酶活性受到抑制后细胞形态学的改变。方法 :以脂质体LipofectAMINE 2 0 0 0介导将硫代磷酸修饰的端粒酶mTR反义寡核苷酸 (ASODN)转染体外分离纯化的 (SD)大鼠精原细胞 ,采用生物发光技术检测精原细胞中端粒酶活性的改变 ,光镜及电镜观测端粒酶活性抑制后A型精原细胞的形态学变化。结果 :mTR ASODN可明显抑制精原细胞端粒酶活性 (P <0 .0 5 )。ASODN转染 4 8～ 72h后 ,光镜下可见细胞生长缓慢 ,染色质浓缩 ,胞质出现颗粒样物质 ,72h后死细胞数较空白对照组明显增多。电镜下可见部分细胞核异染色质浓聚集、浓缩和边集 ,胞质胞核内均可见空泡 ;部分细胞核完全固缩。单纯脂质体组及正义寡核苷酸对照组形态学均无显著变化。结论 :mTR ASODN可明显抑制体外培养的A型精原细胞端粒酶活性 ,并在转染 4 8～ 72h后使细胞形态学出现明显改变     

反义人端粒酶RNA亚基对胆囊癌端粒酶活性及细胞生长的抑制作用

目的探讨反义RNA技术对胆囊癌细胞端粒酶活性的影响及对胆囊癌细胞生长增殖的作用。方法根据测定的胆囊癌人端粒酶RNA亚基(hTR)基因的序列结果，并根据真核表达载体多克隆酶切位点的物理图谱，体外合成反义RNA及正义RNA的基因序列，并构建入pTriEx-4真核表达载体，酶切鉴定正确后，采用脂质转染法导入胆囊癌细胞。TRAP法检测端粒酶活性。流式细胞仪检测细胞周期及凋亡情况，电镜观察细胞微观形态变化。结果实验组胆囊癌细胞的端粒酶活性较对照组明显减低，正义组、转染空载体及单纯脂质体转染的细胞端粒酶活性与未转染细胞比较则变化不明显。反义RNA基因作用后，G0／G1期细胞比例明显增高，S期细胞比例明显降低。细胞的分裂、增殖受到明显抑制。反义RNA基因转化后，10d凋亡率为11．10％。13d后凋亡率为29．02％。电镜下可见明显凋亡细胞。结论反义RNA技术对胆囊癌细胞端粒酶活性有明显的抑制作用；并抑制胆囊癌细胞的生长，促进细胞的凋亡；且克服了反义寡核苷酸作用时间短的缺点。     

反义核酸对大鼠A型精原细胞端粒酶活性及其mTR亚基表达的影响

目的 :探讨小鼠端粒酶RNA(mTR)基因的反义寡核苷酸 (ASODN)对体外培养的大鼠A型精原细胞端粒酶活性及其mTR亚基表达的影响。　方法 :以脂质体LipofectAMINE 2 0 0 0 (LF 2 0 0 0 )介导将硫代磷酸修饰的端粒酶mTR的ASODN、正义寡核苷酸 (SODN)、随机寡核苷酸 (RODN)及单纯脂质体组分别转染体外分离纯化的SD大鼠A型精原细胞 ,采用生物发光技术和TRAP SYBR Green染色法检测精原细胞中端粒酶活性的改变 ,原位杂交检测mTR基因mRNA的表达。　结果 :端粒酶mTR的ASODN明显抑制精原细胞端粒酶活性 (P <0 .0 1) ;mTR ASODN作用于精原细胞 2 4h后 ,mTR基因mRNA的表达水平显著下调。单纯脂质体组及SODN、RODN对照组均无此抑制作用 (P >0 .0 5 )。　结论 :硫代修饰的端粒酶mTR ASODN可使精原细胞端粒酶活性明显受到抑制 ,其机制可能是在mTR基因转录水平影响精原细胞端粒酶活性。     

脂质体介导的聚集素反义寡核苷酸对膀胱癌EJ细胞的作用

目的探讨clusterin对膀胱癌EJ细胞生物学特征的影响. 方法应用脂质体包裹的clusterin反义寡核苷酸转染EJ细胞,阻断其表达,通过3′端末端标记、MTT试验、伊红排斥实验等方法观察clusterin反义基因对EJ细胞增殖、凋亡、坏死等生物学行为的影响. 结果转染clusterin反义寡核苷酸组(反义组)EJ细胞凋亡增加,第5天达峰,凋亡率80%,与转染clusterin正义寡核苷酸组(正义组)比较,差异有显著性意义(P<0.05).反义组细胞死亡率增加,以转染后1～4 d明显(13.8%～33.3%),显著高于相应正义组(P<0.05).反义组细胞增殖活性降低,以转染后第6、7天明显,显著高于相应正义组(P<0.05). 结论 clusterin是重要抗凋亡因子,其表达与膀胱癌细胞的生存、增殖、抗凋亡发展密切相关.     

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.     

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.     

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.