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1.
K-ras基因突变与结直肠癌临床病理因素的关系   总被引:1,自引:0,他引:1  
Yuan Y  Hu HG  Ye XX  Shen H  Zheng S 《中华外科杂志》2010,48(16):1247-1251
目的 建立K-ras基因简单、快捷、经济的检测方法,并将检测结果与临床病理因素进行相关性分析.方法 采用PCR-直接测序法和聚合酶链式反应-限制性片段长度多态性测序法(PCR-RFLP)同时检测40例结直肠癌肿瘤组织标本K-ras基因第12、13密码子突变情况;另113例患者仅用PCR-RFLP-测序法检测K-ras基因第12、13密码子突变情况.将K-ras基因突变检测结果与临床病理因素进行相关性分析.结果 40例结直肠癌患者的肿瘤组织经常规PCR扩增后直接测序无一例发现K-ras基因第12、13密码子的突变;而这40例标本经PCR-RFLP-测序法检测发现:8例含有K-ras基因第12密码子突变,3例含有K-ras基因第13密码子突变,总突变检出率为27.5%(11/40).153例结直肠癌肿瘤组织标本经PCR-RFLP-测序法检测共发现突变58例,突变率为37.9%(58/153),其中第12密码子突变46例,第13密码子突变12例.G→A是K-ras基因突变最常见的突变形式(25/58,43.1%).K-ras基因第12和13密码子突变与患者性别、肿瘤浸润深度、分化程度、与淋巴结转移、远处转移及分期无明显相关性(P>0.05),与年龄、肿瘤发生部位密切相关(P<0.05),年龄越大突变概率越低,突变最常发生的肿瘤部位为升结肠.结论 PCR-RFLP-测序法能够快捷、灵敏地检测出肿瘤组织中K-ras基因的突变,适合作为K-ras基因突变常规检测方法.K-ras基因第12和13密码子突变是结直肠癌中一个常见的分子事件,与患者年龄、肿瘤发生部位有相关性.  相似文献   

2.
目的:探讨结直肠癌中K-ras基因突变状态及其与临床病理特征的关系.方法:应用实时荧光定量PCR( RT-qPCR)和直接基因测序法检测200例结直肠癌患者癌组织K-ras基因突变状态.将两种检测结果进行对比,并结合临床病理资料分析其意义.结果:200例结直肠癌患者中,RT-qPCR检测出突变63例,突变检出率31.5%;经直接基因测序,测序成功的样品169例,检出突变50例,突变检出率29.6%.其中第12密码子GGT→GAT最常见,占34.9%( 22/63);其次是第13密码子GGC→GAC,占28.6%(18/63);第12密码子GGT→CGT最少,全组未见(0/63).两种方法突变检测一致率为98%.K-ras基因突变与肿瘤分化程度有关( P<0.05),而与患者性别、年龄、肿瘤部位、淋巴结转移及TNM分期无明显关系(均P>0.05).结论:RT-qPCR能快速、敏感、准确地检测结直肠癌K-ras基因突变位点,为临床靶向治疗提供可靠的参考依据.  相似文献   

3.
目的 阐明CDKN2/P16基因突变与原发性肝癌发生的关系。方法 采用PCR-SSCP及PCR产物直接银染测序技术,对30例原发性肝癌组织中CDKN2/P16基因第二外显子进行突变检测。结果 检出2例(2/30)CDKN2基因突变,均是位于第124位密码子的错义突变。结论 CDKN2基因的突变虽不频发,但在原发性肝癌的发生中起了一定作用。  相似文献   

4.
ras基因家族包含H-ras、K-ras和N-ras3种,3种ras基因结构相似,表达产物通称p21蛋白,在细胞间的信息传递及细胞分化方面起着关键作用.ras基因在第12、13或61位密码子发生点突变后,p21蛋白的氨基酸序列及空间构象随之发生变化,使得p21蛋白在信息传递后仍不失活,刺激细胞无限制性生长,并认为在某些肿瘤的发生及发展中起着关键性作用.胰腺癌K-ras基因点突变率高达67%~100%,突变几乎总是发生在第12位密码子上,偶报告发生在第13位密码子;突变方式也几乎皆为野生型GGT突变为CGT、GTT及GAT,偶报告突变为TGT等,而正常胰腺组织、急慢性胰腺炎、胰岛素瘤、胰腺良性囊肿及胰腺其他疾病  相似文献   

5.
目的 观察结直肠癌组织中K-ras瑚基因突变,检测患者外周血清CEA水平,探讨两者与结直肠癌肝转移的关系.方法 应用实时荧光定量聚合酶链反应(PCR)方法和基因测序技术检测100例结直肠癌组织中K-ras基因1号外显子12、13密码子突变,化学发光法检测患者血清CEA水平,结合其临床资料分析.结果 100例结直肠癌组织中K-ras基因突变者39例(39.0%),其中12号密码子突变31例,13号密码子突变8例.36例肝转移组中K-ras基因突变23例(23/36,63.9%);64例无肝转移组中K-ras基因突变16例(16/64,25.0%).36例有肝转移患者外周血清CEA水平为(66.79±127.46)μg/L,其中25例(25/36,69.4%)出现不同程度CEA升高;64例无肝转移患者外周血清CEA水平为(4.93±4.62)μg/L,其中24例(24/64,37.5%)CEA升高.两个指标联合检测,肝转移组的阳性率(22/36,61.1%)明显高于无肝转移组的阳性率(5/64,7.8%),P<0.01.其预测结直肠癌肝转移的敏感性及特异性分别为61.1%和92.2%.结论 K-ras基因突变、血清CEA水平与结直肠癌肝脏转移有密切相关,联合检测癌组织K-ras基因突变和血清CEA水平可预测结直肠癌肝转移.  相似文献   

6.
目的探讨新疆地区维吾尔族和汉族直肠癌患者K-ras基因外显子2中第12、13位密码子的突变情况,为新疆地区直肠癌患者的临床及基因诊断提供理论参考。方法收集2010年1月至2011年12月期间在新疆维吾尔自治区人民医院手术切除的临床资料完整的直肠腺癌石蜡包埋组织144例,同时选取距癌缘近端10 cm以远的正常直肠石蜡包埋组织144例作为对照组。提取癌组织DNA,经聚合酶链反应扩增K-ras基因,采用DNA直接测序法检测K-ras基因的突变情况。结果 144例直肠癌患者中有32例(22.22%)K-ras基因第12、13位密码子发生突变,汉族与维吾尔族突变率分别为20.41%(20/98)和26.09%(12/46),二者比较差异无统计学意义(P>0.05)。对照组中均未见K-ras基因第12、13位密码子的突变。K-ras基因突变仅与肿瘤浸润深度有关(T1+T2为25.0%,T3+T4为75.0%,P=0.01),与民族、性别、肿瘤部位、分化程度及淋巴结转移均无关(P>0.05)。结论在维吾尔族与汉族直肠癌患者中K-ras基因第12、13位密码子突变均很常见,但在突变频率上维吾尔族与汉族直肠癌患者间无差异,K-ras基因突变与直肠癌的侵犯深度有关。  相似文献   

7.
目的 研究K-ras基因在人胆管癌中的突变。方法 采用聚合酶链式反应-限制性片段长度多态性(Polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP)检测了43例胆管癌17例胆管良性病变K-ras基因12密码子点突变情况。结果 胆管癌K-ras基因12密码子点突变率为81.4%(35/43)。1例胆总管囊肿中检测到突变。结论 K-ras基因12密码子点突变在胆管癌发生中起重要作用,其检测有可能用于临床进行胆管癌的基因诊断。  相似文献   

8.
目的观察结直肠癌组织中K-ras基因突变情况,术前检测患者血清CEA水平,探讨两者与结直肠癌临床病理的关系。方法应用实时荧光定量PCR方法和基因测序技术检测100例结直肠癌组织中K-ras基因12、13密码子突变情况,化学发光法检测患者血清CEA水平,结合其临床病理资料进行统计学分析。结果 K-ras基因突变者39例(39.0%),其中12号密码子突变31例,13号密码子突变8例。有淋巴结转移者K-ras基因突变率(57.8%)明显高于无淋巴结转移者(23.6%),有肝脏转移者K-ras基因突变率(62.5%)明显高于无肝转移者(34.5%),TNM分期Ⅲ、Ⅳ期K-ras基因突变率(56.5%)明显高于Ⅰ、Ⅱ期(24.1%)。K-ras基因突变率与肿瘤大小、部位、肿瘤浸润深度、分化程度无密切关系。49例CEA水平超出正常范围(49.0%)。有淋巴结转移或肝转移患者CEA水平显著高于无淋巴结或肝转移患者(P0.05)。在肿瘤不同的临床病理分期(Duke's、TNM分期)间CEA水平差异有统计学意义,Duke's D期和TNMⅢ、Ⅳ期出现高阳性率。CEA水平在肿瘤不同浸润深度、体积及分化程度上无明显差异。结论癌组织K-ras基因突变和血清CEA水平超出正常水平预示结直肠癌可能合并淋巴结转移或肝转移,是预后不良的指标。  相似文献   

9.
目的 观察毒性多结节性甲状腺肿(TMG)促甲状腺激素受体(TSHR)基因突变情况及探讨TSHR基因突变引起TMG细胞的生物学行为的改变.方法 将21例TMG患者手术切除标本中甲状腺结节组织及其作为对照组的结节周围正常的甲状腺组织分别提取DNA,对目的 基因片段进行扩增聚合酶链反应(PCR)反应及DNA测序分析.结果 21例TMG标本中发现11例存在TSHR基因突变(52.3%).其中,4例碱基插入性突变:在1928和1929位核酸之间播入了一个鸟嘌呤(G),使该密码子609位以后的氨基酸发生了移码突变;7例为点突变:第613位密码子的第一碱基即第1937位核苷酸发生T→A转换,使密码子由TAC变为AAC,酪氨酸被天冬酰胺置换即Tyr613Asn(TAC→AAC).对照组未发现TSHR基因突变.结论 基因突变的TSHR对TMG细胞的生物学行为的改变起着一定作用.  相似文献   

10.
甲状腺癌的分子生物学研究进展   总被引:4,自引:1,他引:3  
甲状腺癌是多因素起源的癌基因和抑癌基因相互失衡导致的疾病。随着在分子和蛋白水平上对肿瘤研究的进展,现已发现不少癌基因和肿瘤标志物(tumormakers)与甲状腺肿瘤的发生、发展、转归有关,将为其病因、诊断、预测和基因治疗带来前景。一、甲状腺癌相关癌基因1.ras基因:60%的K-ras基因在放射性相关的甲状腺肿瘤中被激活。缺碘地区的甲状腺乳头状腺癌其ras基因突变率比非缺碘地区低。H-ras和N-ras的61密码子突变也见于多种甲状腺癌中。多数研究认为ras基因突变在甲状腺癌发生的早期起作用,但对细胞恶性变则并不起决定性作用。2.HER-2/neu…  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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