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1.
目的 探讨腺病毒载体介导的胞嘧啶脱氨酶(CD)基因转染和CD/5-FC对直肠癌细胞的杀伤作用。方法 用重组腺病毒介导外源CD基因转移到人直肠癌细胞株HR-8348,通过检测腺病毒的转导效率,CD基因表达,以及集落形成实验,细胞存活率测定,裸鼠移植癌治疗实验,观察分析CD/5-FC对癌细胞的杀伤作用,结果 重组腺病毒介导CD基因转染,在癌细胞中得到高效表达。CD/5-FC系统对转染含CD重组腺病毒HR-8348细胞的集落形成,细胞生长均有明显的抑制作用,而对无CD基因转染癌细胞无影响,在转染和未转染CD基因的HR-8348混合体系中,CD/5-FC除了杀伤转染CD基因的癌细胞外,对周围的无CD转染癌细胞也有明显的杀伤作用。表现出很强的“旁观者效应”。裸鼠皮下移植癌治疗实验结果表明,CD/5-FC对HR-8348实体癌生长抑制率为71.5%。结论 CD/5-FC对腺病毒介导CD基因转染的直肠癌细胞有很强的杀伤作用和显著的“旁观者效应”。  相似文献   

2.
目的观察逆转录病毒介导的双自杀基因对肝内胆管癌细胞HCCC-9810的杀伤作用,探讨肝内胆管癌的基因治疗方法。方法在脂质体的介导下将含有双自杀基因的逆转录病毒载体PwzlneoCDglytk导人包装细胞PA317,经G418筛选后大量培养产病毒的阳性克隆PA317/CD tk细胞株,收集病毒上清,转染HCCC-9810细胞,再次经G418筛选,获得稳定表达双自杀基因的HCCC-9810/CD tk细胞株。用RT-PCR检测双自杀基因的表达。给予前体药物5-氟胞嘧啶(5-flourocytosine,5-FC)和(或)无环鸟苷(ganciciovir,GCV)后,用MTT法测定转基因组及未转基因组HCCC-9810细胞的存活率。结果双自杀基因在HCCC-9810细胞中可稳定表达,联合使用5-FC和GCV对细胞增殖的杀伤作用及旁杀伤效应高于单独使用5-FC或GCV。结论逆转录病毒介导自杀基因可有效杀死肝内胆管癌细胞,双自杀基因较单一自杀基因具有更强的抗肿瘤作用。  相似文献   

3.
提高转基因效率,建立高效自杀基因杀伤胃癌肿瘤细胞的方法。方法:首先构建含CD基因的重组腺病毒载体并对载体进行包装、收获病毒上清并纯化。将重组病毒液转染MFC胃癌细胞进行体外抑瘤实验。结果:含CD基因的重组腺病毒载体经酶切鉴定选出:包装纯化后,检测病毒浓度为1.2×1012颗粒/ml。体外实验中,转CD基因的MFC细胞在5-FC作用下生长明显受抑(P<0.01,n=9)。结论:腺病毒载体转导的CD基因提高了转染效率,为大量杀伤胃癌细胞奠定了基础。  相似文献   

4.
目的观察双自杀基因胞嘧啶脱氨酶基因(CD)和单纯疱疹病毒胸苷激酶基因(HSV-tk)对胆管癌细胞的抑制作用以及转染缝隙连接蛋白43(Cx43)基因对旁观者效应的影响。方法四甲基偶氮唑蓝(MTT)法测定双自杀基因系统作用下和/或Cx43基因转染后胆管癌QBC939细胞的细胞抑制率及旁观者效应。RT-PCR检测双自杀基因和Cx43基因的表达。Westernblot检测Cx43蛋白的表达。结果CD和tk基因在QBC939/CD tk细胞和QBC939/CD tk Cx43细胞中获得稳定表达。mRNA和蛋白水平均显示转染Cx43基因的QBC939/CD tk Cx43细胞的Cx43的表达量明显增加。双自杀基因系统对QBC939/CD tk Cx43细胞组的抑制作用较QBC939/CD tk细胞组高。结论双自杀基因系统对胆管癌细胞有明显的抑制作用,Cx43基因的导入可明显增强旁观者效应,提高对肿瘤细胞的抑制作用。  相似文献   

5.
目的观察胞嘧啶脱氨酶基因(CD)和单纯疱疹病毒胸苷激酶基因(HSV-tk)双杀基因对胆管癌细胞和裸鼠胆管癌移植瘤的抑制作用.方法在脂质体介导下将质粒pWZLneoCDglytk转入胆管癌QBC939细胞,经G41 8筛选获得稳定表达双自杀基因的QBC939/CD+tk细胞株,RT-PCR法鉴定CD和tk基因的表达.体外给予前体药物5-氟胞嘧啶(5-Fc)和/或丙氧鸟苷(GCV),作用于QBC939/CD+tk细胞,MTT法测定细胞抑制率.裸鼠皮下注射QBC939/CD+tk细胞,成瘤后腹腔注射前体药物,用药前后测量移植瘤的大小判断疗效.结果CD和tk基因在QBC939/CD+tk细胞中获得稳定表达.联合5-Fc和GCV与任何单一药物相比,在较小剂量下可产生较强的抑制作用.随着药物浓度的提高,对细胞生长的抑制作用也相应地提高.前体药物对QBC939/CD+tk细胞早期生成的移植瘤抑制作用明显,肿瘤可完全消失,并观察到明显的局部旁观者效应.结论双自杀基因系统在体内外对胆管癌细胞和裸鼠移植瘤均有明显的抑制效果,旁观者效应也很明显.  相似文献   

6.
体外腺病毒介导的HSV-tk/GCV对产生AFP的人肝癌细胞的影响   总被引:1,自引:0,他引:1  
目的 观察腺病毒介导的单纯疱疹病毒胸苷激酶基因(HSV-tk)/丙氧鸟苷(GCV)自杀基因系统在体外对人肝癌细胞的选择性杀伤效应。方法 采用含有甲胎蛋白基因启动子、增强子和HSV-tk基因的嵌合基因,插入腺病毒中形成重组腺病毒(AdrAFPTK),将该重组腺病毒分别感染体外培养的甲胎蛋白阳性人肝癌细胞BEL-7402和甲胎蛋白阴性人肝癌细胞SMMC-7721,逆转录聚合酶链式反应(RT-PCR)检测HSV-tk基因的转录表达,观察GCV对人肝癌细胞的选择性杀伤作用。结果GCV在体外对重组腺病毒转染的甲胎蛋白阳性的人肝癌细胞BEL-7402有明显的杀伤作用和“旁观者效应”,而对重组腺病毒转染的甲胎蛋白阴性的人肝癌细胞SMMC-7721无明显作用。结论 在体外,表达HSV-tk基因的甲胎蛋白的人肝癌细胞可被受甲胎蛋白基因表达调控序列控制的自杀基因HSV-tk特异性杀伤,表现出极高的细胞专一性。重组腺病毒AdrAFPTK可望用于肝癌的特异性基因治疗。  相似文献   

7.
腺病毒介导HSV-TK/CD基因对胆管癌体内体外的杀伤作用   总被引:1,自引:3,他引:1  
目的 研究HSV TK /CD融合基因对胆管癌的杀伤作用。方法 腺病毒介导HSV TK /CD融合基因体外转染胆管癌细胞QBC93 9,与单自杀基因对照 ,四甲基偶氮唑蓝 (MTT )比色法测定肿瘤细胞生长抑制率 ,旁观者效应 ;构建胆管癌裸鼠模型 ,瘤内注射重组腺病毒 ,观察肿瘤生长情况。结果 与单一自杀基因转染组对照 ,双基因组表现出更强的抗肿瘤作用。当给予相同浓度的前体药物时 ,融合基因组 ,CD组和TK组的肿瘤细胞存活率分别为 3 .1%、3 2 .1%、5 5 .2 %。体内实验显示 ,融合基因组的肿瘤生长受到明显抑制 ,抑制率达 70 .7%。单自杀基因TK、CD组则分别为 41.2 %和 5 5 .7% ,差异有统计学意义 (P <0 .0 5 )。组织学检查可见实验组肿瘤发生明显坏死 ,这种现象在融合基因治疗组尤为明显。结论 与单自杀基因相比 ,HSV TK/CD融合基因在体内和体外都具有更强的抗肿瘤活性。  相似文献   

8.
目的探讨含胞嘧啶脱氨酶(CD)/5-氟胞嘧啶(5-Fc)单纯疱疹病毒1型胸腺嘧啶激酶基因融合自杀基因(CD/TK)的重组腺病毒治疗肝癌的疗效。方法重组腺病毒Ad.CD/TK感染人肝癌细胞Bel7402,用噻唑蓝(MTT)方法观察不同浓度5-Fc和无环鸟苷(GCV)对其杀伤效应; 建立Bel7402裸鼠皮下移植瘤模型,瘤内注射Ad.CD/TK,腹腔注射GCV(50 mg/kg体重)和/或5- Fc(500 mg/kg体重)10 d,观察肿瘤生长抑制效应。结果在对感染Ad.CD/TK的Bel7402细胞的体外杀伤实验中,GCV的IC50为(12.1±2.3)μmol/L,5-Fc的IC50为(223±15)μmol/L,并且两者有协同效应。在Bel7402裸鼠移植瘤模型中,联合Ad.CD/TK和GCV、5-Fc能够显著抑制肿瘤的生长。结论腺病毒为载体的双自杀基因转染和前药的应用对人肝癌细胞的体内、体外治疗疗效明显。  相似文献   

9.
目的 观察腺病毒介导的单纯疱疹病毒胸苷激酶基因(HSV-tk)/丙氧鸟苷(GCV)自杀基因系统在体外对人肝癌细胞的选择性杀伤效应。方法 采用含有甲胎蛋白基因启动子,增强子和HSV-tk基因的嵌合基因,插入腺病毒中形成重组腺病毒(AdrAFPTK),将该重组腺病毒分别感染体外培养的甲胎蛋白阳性人肝癌细胞BEL-7402和甲胎蛋白阴性人肝癌细胞SMMC-7721,逆转录聚合酶链式反应(RT-PCR)检测HSV-tk基因的转录表达,观察GCV对人肝癌细胞的选择性杀伤作用。结果 GCV在体外对重组腺病毒转染的甲胎蛋白阳性的人肝癌细胞BEL-7402有明显的杀伤作用和“旁观效应”,而对重组腺病毒转染的甲胎蛋白阴性的人肝癌细胞SMMC-7721无明显作用。结论 在体外,表达HSV-tk基因的甲胎蛋白的人肝癌细胞可被受甲胎蛋白基因表达调控序列控制的自杀基因HSV-tk特异性杀伤,表现出极高的细胞专一性,重组腺病毒AdrAFPTK可望用于肝癌的特异性基因治疗。  相似文献   

10.
目的:观察逆转录病毒介导的双自杀基因CD、HSV-tk共表达对肝门部胆管癌细胞FRH的杀伤作用,探讨肝门部胆管癌的基因治疗方法.方法:在脂全Lipofectamine的介导下将含有双自杀基因的逆转录病毒载体PwzheoCDglytk转导入包装细胞PA317,经G418筛选后培养产病毒的阳性克隆PA317/CD 细胞株,收集其病毒上清,转染FRH细胞,再次经G418筛选,获得稳定表达双自杀基因的FRH/CD tk细胞株,用RT-PCR检测双自杀基因的表达.给予前体药物5-氟胞嘧啶(5-flourocy-tosine,5-Fc)或无环鸟苷(Gancicioir,GCV)后,用MTT法测定转基因组及未转基因组FRH细胞的存活率.结果:双自杀基因在FR细胞中可稳定表达,联合使合5-Fc和GCV对靶细胞增殖的杀伤作用及旁杀伤效应高于单独使用5-Fe或GCV.结论:逆转录病毒介导自杀基因可有交往杀死肝门部胆管癌细胞.双自杀基因共表达较单一自杀基因有更强的抗瘤作用.  相似文献   

11.
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 (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.  相似文献   

12.
Don Dame 《Artificial organs》1996,20(5):613-617
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.  相似文献   

13.
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.  相似文献   

14.
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.  相似文献   

15.
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.  相似文献   

16.
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.  相似文献   

17.
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.  相似文献   

18.
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.  相似文献   

19.
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.  相似文献   

20.
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.  相似文献   

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