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1.
目的探讨以SSX-1基因mRNA为特异性标记物检测肝细胞肝癌(HCC)患者外周血中肿瘤细胞的可行性及其临床意义。方法用逆转录-聚合酶链反应(RT-PCR)方法对25例HCC患者及20例非肝癌患者的外周血进行SSX-1 mRNA检测。用同样方法检测HCC组织、癌旁组织、硬变肝组织及正常肝组织中SSX1 mRNA的表达。为验证结果的可靠性,对RT-PCR产物进行DNA测序。结果SSX-1 mRNA在HCC组织及其外周血中的表达阳性率分别为60%(15/25)和40%(10/25)。癌旁组织、硬变肝组织及正常肝组织中均未发现该基因的表达。测序结果表明所得到的cDNA确为SSX-1基因。SSX-1基因的表达与HCC患者的年龄、性别、肿瘤大小、TNM分期、分化程度、血清AFP水平、肝炎病毒感染及短期(半年)复发临床指标无相关性(P〉0.05)。33%(3/9)AFP正常(〈20μg/L)的HCC患者外周血中有SSX-1基因表达。结论SSX-1 mRNA在HCC外周血中高特异性表达,可作为肿瘤特异性标记物用于检测HCC患者外周血中的肝癌细胞,具有辅助定性诊断价值;部分AFP正常(〈20μg/L)HCC患者的外周血中有SSX-1 mRNA的表达,提示两者联合应用有助于提高HCC的检出率。  相似文献   

2.
目的 探讨肿瘤-睾丸抗原9(CT9)基因mRNA在肝细胞癌中的表达情况。方法 用逆转录-聚合酶链反应(RT—PCR)方法对45例肝细胞癌患者癌组织和相应癌旁组织中CT9基因mRNA的表达情况进行检测,并对其中3例RT—PCR扩增产物的目的片段进行DNA序列测定。结果 45例肝细胞癌患者中,23例表达CT9mRNA,表达阳性率为51.1%,相应的癌旁组织中则均为阴性;3例DNA测序结果表明RT—PCR产物确为CT9 cDNA。CT9 mRNA的表达与患者年龄、性别、肿瘤大小、分化程度、血清AFP水平、HBV和HCV感染无相关性(P〉0.05)。结论 CT9 mRNA在肝细胞癌中呈高比例、特异性表达,可望成为肝细胞癌免疫治疗的理想靶位。  相似文献   

3.
目的探讨原发性肝癌癌旁组织生长激素受体(GHR)的基因表达情况或变化。方法采用放射配体法、半定量的逆转录-多聚酶链式反应(RT—PCR)对45例原发性肝癌(HCC)的癌旁组织进行了生长激素受体(GHR)的检测,以8例正常肝组织作为对照,并随机选择12例RT—PCR阳性扩增产物直接进行DNA序列测定。结果在蛋白表达水平,应用放射性配体法于正常肝组织与45例HCC癌旁组织中检测到单一的GH特异性结合位点(即GHR)。对照组肝组织GHR的RT值为40.2932±3。5667.fmol/mg;protein,Kd为0.6167±0.1007nmol/L。HCC癌旁组织GHR的RT为33.6283±3.6218fmol/mg。protein,Kd值为0.6319±0.1978nmol/L,与正常肝组织相比,HCC癌旁GHR的RT降低(P〈0.05).而Kd无显著性改变(P〉0.05)。半定量的RT—PCR法发现GHRmRNA在正常肝组织与HCC癌旁组织的表达率均为100%。GHRmRNA在正常肝组织与原发性肝癌癌旁组织的表达两者之间差异无显著性(P〉0.05)。结论全部肝癌癌旁组织在蛋白与mRNA水平均表达GHR,在其受体功能未明的情况下,目前rhGH在肝癌患者中的使用应慎重,以免造成肿瘤的增殖与复发。  相似文献   

4.
以SSX基因mRNA作为肿瘤标志物检测循环中的肝癌细胞   总被引:1,自引:0,他引:1  
目的 探讨检测肝细胞癌 (HCC)患者外周血中SSX基因mRNA的临床意义。方法 以简易“降落”PCR(stepdownRT PCR)方法检测HCC患者外周血中SSX基因mRNA的表达 ,并对SSX基因的表达与临床指标和预后的关系进行了分析。结果  4 2例肝癌患者肝癌组织中SSX基因mRNA阳性率为 85 7% (36 4 2 ) ,而癌旁组织、肝硬化组织和正常肝组织均未发现有SSX基因的表达。外周血标本中 ,有 17例可检测到SSX基因的表达 ,占 4 0 5 %。肝癌组织不表达SSX基因者以及健康志愿者的外周血中未检测到其表达。外周血中SSX基因的表达与年龄、性别、肿瘤分化程度、TNM分期、肿瘤大小、血清AFP水平和HBV感染无显著相关性 (P >0 0 5 )。除肝组织不表达SSX和死于并发症的 7例患者外 ,在随访 7 7± 5 4月的过程中 ,16例外周血中表达SSX基因mRNA者有 8例出现了复发和 或转移 ,其中 7例已经死亡 ,另 1例带瘤生存 ,而 19例阴性者仅 2例出现了复发和 或转移 ,二者相比差异有显著性 (P =0 0 2 8)。结论 外周血中SSX基因表达与复发、转移密切相关 ,SSX基因可作为检测HCC患者循环肿瘤细胞的标志基因。  相似文献   

5.
以MAGE基因mRNA为特异标记检测肝癌患者外周血肿瘤细胞   总被引:5,自引:0,他引:5  
目的 以黑色素瘤抗原-1(MAGE-1)和MAGE-3基因mRNA为特异性标记物,检测肝细胞癌(HCC)患者外周血中的肿瘤细胞,并探讨其临床意义。方法 采集25例HCC患者及20例健康志愿者的外周血,用巢式RT-PCR方法检测外周血单核细胞(PBMC)中MAGE-1和MAGE-3基因mRNA。同时用RT-PCR方法检测HCC患者肝癌组织中上述MAGE基因的表达。结果 44%(11/25)和36%(9/25)HCC患者的PBMC中可分别检测到MAGE-1和MAGE-3基因mRNA,而相应肝癌组织中MAGE-1和MAGE-3的表达率分别为68%(17/25)和56%(14/25);在64%(16/25)的HCC患者PBMC中至少可检测到一种MAGE基因mRNA;肝癌组织中不表达MAGE基因的病例以及20例健康志愿者的PBMC中均未测出MAGE基因mRNA。HCC患者PBMC中两种MAGE基因mRNA的检出率与肿瘤TNM分期、直径密切相关,而与肿瘤分化程度、血清AFP水平无相关性。但9例血清AFP正常或轻度升高者(<50ng/ml),6例的PBMC中MAGE-1和/或MAGE-3 mRNA为阳性。结论 MAGE-1和MAGE-3基因mRNA可作为特异性肿瘤标记物用于检测HCC患者PBMC中的肝癌细胞,巢式RT-PCR的敏感性及特异性较高,其检测结果可能有助于判断HCC患者的预后。  相似文献   

6.
目的检测CT10基因mRNA在肝细胞肝癌 (HCC)中的表达情况 ,了解肿瘤睾丸抗原CT10基因mRNA在HCC中的表达是否具有肿瘤特异性 ;对CT10HLA A2限制性CTL表位进行预测。方法用逆转录 聚合酶链反应 (RT PCR)方法对 45例HCC患者癌组织和相应癌旁组织的CT10基因mRNA表达情况进行了检测 ,对其中 3例RT PCR扩增产物的目的片段进行DNA序列测定 ;利用超基序法和量化基序法联合预测CT10HLA A2限制性CTL表位。结果 45例HCC患者中 ,19例表达CT10mRNA ,阳性率为 42 % ,相应的癌旁组织中结果均显示阴性 ;3例DNA测序结果表明RT PCR产物确为CT10cDNA。CT10的表达与年龄、性别、肿瘤体积、分化程度、血清甲胎球蛋白 (AFP)水平、HBV和HCV感染无显著相关性 (P >0 0 5 ) ;预测发现 9个HLA A2限制性CTL表位。结论CT10在HCC中呈高比例、特异性表达 ,可望成为HCC免疫治疗的理想靶位。  相似文献   

7.
CT9和CT10基因在肝细胞癌中的表达及临床意义   总被引:1,自引:0,他引:1  
目的检测肿瘤睾丸抗原CT9和CT10基因mRNA在肝细胞癌(HCC)中的表达情况。方法用逆转录-聚合酶链反应(RT-PCR)方法对HCC病人的癌组织和相应癌旁组织及对照(肝硬化和正常肝组织)中CT9和CT10mRNA的表达进行了检测,随机选取每一种抗原RT-PCR呈阳性的产物4例进行DNA序列测定,结合AFP、抗HCV、HgsAg、肿瘤直径、分化程度等临床指标进行分析。结果56例HCC病人中,CY9、CT10基因mRNA阳性率分别是51.8%(29/56)和44.6%(25/56),至少表达一种CTA者达66.1%(37/56),表达两种者为30.4%(17/56);电泳显示PCR产物与阳性对照大小一致;而癌旁组织中仅有CT9基因nRNA表达为阳性,阳性率为7.1%(4/56),但表达强度明显弱于在癌组织中的表达,进一步的病理检查未发现CY9表达呈阳性的癌旁组织中有癌细胞。所测10例肝硬化和10例正常肝组织均未检测到CT9和CT10的表达。DNA测序结果表明RT-PCR产物确为CT9和CT10cDNA。CT9和CT10的表达与年龄、性别、肿瘤大小、分化程度、AFP水平、HBV和HCV感染无显著相关性(P〉O.05)。而在部分AFP正常(〈25ng/L)HCC病人中存在CT9和CT10基因的表达。结论CT9和CT10基因在HCC中呈高比例、高特异表达,为HCC免疫治疗提供了新的理想靶位;CT9和CT10同时在HCC中表达,为应用以这些肿瘤抗原为基础的多价瘤苗治疗HCC提供了实验依据。  相似文献   

8.
中期因子mRNA在原发性肝癌中的表达及其意义   总被引:3,自引:1,他引:2  
目的:探讨中期因子(MK)mRNA在原发性肝细胞癌(HCC)中的表达及其意义。方法:收集33例人肝细胞癌组织、配对癌旁肝组织及10例肝良性肿瘤组织、5例正常肝组织、采用TRIzol一步抽提法提取总RNA,应用逆转录-聚合酶链反应(RT-PCR)检测MKmRNA的表达。结果:28例肝细胞癌可见MKmRNA表达,阳性率为84.8%,而配对的癌旁组织中仅见3例表达,正常肝脏和肝脏良性肿瘤细胞无1例阳性。统计学分析表明,肝细胞癌肝细胞癌MKmRNA表达率与其他各组的差异有非常显著性(P<0.01),但肝细胞癌MKmRNA表达与其组织学类型、病理分级、瘤体大小、包膜状况及其甲胎蛋白(AFP)值之间差异无显著性(P>0.05)。结论:肝细胞癌在mRNA水平上表达MK增加,检测MK表达情况可作为HCC的辅助诊断指标之一,对术后制定进一步治疗方案也有参考价值。  相似文献   

9.
目的 探讨C-myc基因扩增、p16基因变异和乙型肝炎病毒(HBV)DNA整合与肝细胞癌(HCC)的相关性。方法 应用差异聚合酶链反应(d-PCR)分析C-myc基因扩增,用聚合酶链反应(PCR)和单链构象多态(SSCP)分析p16基因变异,用PCR法检测整合型HBV-DNA。结论 (1)HCC组外周血、癌和癌旁肝组织中C-myc基因扩增阳性率分别为48%(14/29)、45%(13/29)和52%(15/29),3者间差异无显著意义(x^2=0.0008,P>0.05,自由度ν=2),但明显高于肝硬化(LC)组外周血和LC组织阳性率[0(0/12)和8%(1/12),P均小于0.05]。(2)29例癌组织中有3例p16基因纯合性缺失,未发现点突变。(3)正常肝、肝硬化和肝癌组织中整合型HBV-DNA阳性率分别为14/5(2/14)、67%(8/12)和97%(28/29),3者间差异有显著意义(x^2=29.4345,P<0.01)。结论(1)HCC的发生与C-myc基因扩增密切相关;(2)HCC中p16基因的纯合性缺失发生频率为10%;(3)HBV-DNA整合与HCC的发生密切相关;(4)对外周血的检测能反映肝细胞(C-myc基因扩增和HBV-DNA整合情况。  相似文献   

10.
目的探讨肝细胞癌(HCC)中SFRP1和APC基因启动子甲基化状态及其mRNA表达的关系。方法应用甲基化特异性聚合酶链反应(MSP)和逆转录-聚合酶链反应(RT—PCR)技术,检测30例肝细胞癌(HCC组)及相应的癌旁组织(癌旁组)和10例正常肝组织(正常对照组)中SFRP1及APC基因启动子甲基化状态和mRNA的表达水平,分析甲基化与某些临床参数与mRNA表达的关系。结果HCC组、癌旁组及正常对照组中的SFRP1和APC基因启动子甲基化率分别是11/30,4/30,0/10和14/30,5/30,0/10;HCC组明显高于其余两组(P〈0.05)。SFRP1基因启动子甲基化与临床资料无关(P〉0.05);APC基因启动子甲基化与年龄(〈60)岁和癌肿无假包膜有关(P〈0.05)。HCC组SFRP1基因mRNA表达明显低于其余两组(P〈0.05),各组APC基因mRNA表达差异无显著性。两基因甲基化之间无相关性。结论SFRP1和APC基因启动子甲基化与HCC的形成和进展有关,但HCC组织中基因甲基化与mRNA表达的关系尚不明确。  相似文献   

11.
Background: The duration of action of muscle relaxants is poorly correlated to the rate of decay of their plasma concentration. The plasma concentration of mivacurium may rapidly decrease below its active concentration because of the extensive hydrolysis of mivacurium. By inflating a tourniquet on one upper limb for 3 min after the administration of atracurium, mivacurium or vecuronium, we studied the influence of the initial decline of their plasma concentration on their effect. Methods: In 50 patients anaesthetised with thiopental, isoflurane and fentanyl, the effect of bolus doses of 0.15 or 0.25 mg . kg?1 mivacurium (MIV 15, MIV 25), 0.3 or 0.5 mg . kg?1 atracurium (ATR 30, ATR 50) and 0.06 or 0.1 mg . kg?1 vecuronium (VEC 06, VEC 10) were measured on both arms (evoked response of the adductor pollicis to train-of-four stimulation every 12 s), a tourniquet being applied on one arm just before and during 3 min after the muscle relaxant bolus. Results: Tourniquet inflation of 3 min almost abolished the neuromuscular effect of mivacurium. In the vecuronium groups and in the ATR 50 group, tourniquet inflation did not modify the maximum degree of depression of the twitch response. Also, the duration of action of vecuronium was unaffected by the tourniquet. In the ATR 30 group, times to return of the twitch response to 25% (duration 25%) and 75% (duration 75%) of control response were significantly shorter in the cuffed arm, 23 min vs 27 min, and 41 min vs 45 min, respectively. In the ATR 50 group, only duration 25% was significantly shorter in the cuffed arm (41 min vs 45 min). Conclusion: The results suggest that the rate of decline of the plasma concentration of mivacurium is so rapid, that a very low and almost clinically ineffective concentration is present as soon as 3 min after its administration. The results also indicate that the recovery from a mivacurium-induced neuromuscular blockade is not influenced by the rate of decay of its plasma concentration in patients with genotypically normal plasma cholinesterase.  相似文献   

12.
Abstract: Membrane processes play a pivotal and enabling role in modern replacement therapy for acute and chronic organ failure and in the management of immunologic diseases. In fact, virtually all contemporary extracorporeal blood purification methods employ membrane devices, and the next generation of artificial organs and tissue engineering therapies are almost certain to be similarly grounded in membrane technology. In this short essay, we comment on the similarities and differences among synthetic membranes and their natural counterparts and also provide a critical overview of the demographics and technology of hemodialysis, hemofiltration, apheresis, oxygenation, and emerging membrane technologies and applications.  相似文献   

13.
Abstract Immunoadsorption (1A) therapy with tryptophan (TR-350) or phenylalanine (PH-350) adsorbents has been used to reduce the concentration of serum antibodies in human lymphocyte antigen (HLA)-immunized patients. Other forms of plasma purification have been reported to reduce the level of fibrinogen, which affects the blood properties. In this study we investigated the effects of IA therapy using both adsorbents on plasma fibrinogen and immunoglobulins G and M in 13 patients (8 patients were treated with TR-350, and 5 patients were treated with PH-350). During each session 1 plasma volume (2.8 ± 0.4 L of plasma) was processed through the immunocolumn and then returned to the patient together with the blood cells. Compared with the pretreatment values, the plasma fibrinogen, IgG, and IgM concentrations were significantly reduced after IA therapy (p < 0.01 for TR-350; p < 0.04 for PH-350). There was a positive correlation between the degree of reduction of plasma proteins and the number of IA treatments given. A nonpara-metric test (Wilcoxon's signed-rank test or the Mann-Whitney test) was used for statistical analysis. We conclude from our study that IA therapy effectively lowers the plasma levels of fibrinogen, IgG, and IgM and thus can be considered a valuable alternative to other blood purification methods.  相似文献   

14.
Abstract: Numerous articles have been published on the multiple use of dialyzers and on the effect of different reprocessing chemicals and techniques on the dialyzer biocompatibility and performance. The results often appear contradictory, especially those comparing standard biocompatibility parameters. Despite this confusion, a discerning review of the published works allows certain limited conclusions to be drawn. Reprocessing of used hemodialyzers changes the biocompatibility profile of a dialyzer as defined by the parameters complement activation. leukopenia, and cytokine release. The effect of reprocessing depends on the chemicals and reprocessing technique applied and also on the type of membrane polymer being subjected to the reprocessing procedure. Reports of pyrogenic reactions indicate that the flux of the membrane also influences how suitable it is for safe reuse. An increased risk of allergic and pyrogenic reactions appears to be associated with dialyzer reuse. Furthermore, there has been a lack of investigations into the immunologic effect of the layer of adsorbed and chemically altered proteins that remains on the inner surface of reprocessed dialyzers. We conclude that the clinical benefit of dialyzer reuse cannot be generally accepted from a biocompatibility point of view.  相似文献   

15.
Background : Ketamine in sub-dissociative doses has been shown to have analgesic and phantom-Limb pain, where conventional treatment has often failed. Chronic ischemic pain due to lower extremity arteriosclerosis obliterans often responds poorly to analgesics, and the pain-generating mechanisms are not well understood.
Methods : Eight patients with rest pain in the lower extremity due to arteriosclerosis obliterans were given sub-dissociative doses of 0.15, 0.30, or 0.45 mg/kg racemic ketamine and morphine 10 mg as a 5-min infusion on four separate days in a cross-over, double-blind, randomised protocol. Plasma levels of (S)- and (R)-ketamine and their nor-metabolites were analysed with an enantioselective high-performance liquid chromatography (HPLC) method. Pain levels were evaluated with a visual analogue scale (VAS).
Results : Individual pain levels were highly variable during and after all the infusions but the pooled pain levels showed a dose-dependent analgesic effect of ketamine with a transient but complete pain relief in all patients at the highest dose (0.45 mg/ kg). Side-effects, mainly disturbed cognition and perception, were pronounced and dose-dependent. Morphine 10 mg had an analgesic peak at 20 min and 5/8 patients had complete pain relief. The remaining 3 patients also had high baseline pain scores, indicating a higher analgesic potency for the 0.30 and 0.45 mg/ kg ketamine doses than for morphine 10 mg.
Conclusion : We have demonstrated a potent dose-dependent analgesic effect of racemic ketamine in clinical ischemic pain. Due to a narrow therapeutic window, this analgesic effect is probably best utilised in combination with other analgesics.  相似文献   

16.
Background : It is unclear whether activation of the inducible nitric oxide synthase (iNOS) increases or decreases the extravasation of plasma.
Methods : Chloralose anaesthetised male Wistar rats received E. coli lipopolysacharide (LPS), 3 mg kg-1 i.v., or the corresponding volume of saline, 3 or 5 h before the end of the experiment. Mean arterial pressure (MAP) and heart rate (HR) were recorded. Tissue clearance of radio-labelled albumin, during the last 2 h of each experiment, was determined by a double-isotope method. In separate animals, the serum concentration of nitrite and nitrate was determined, 5 h after LPS or the solvent.
Main Results : LPS initially decreased MAP and lastingly increased HR. In the 3-h LPS animals (n=8), tissue plasma clearance was lower in the heart and calf muscle and increased only in diaphragm, compared to corresponding control animals (n=8). In the 5-h LPS rats, clearance was lowered (n=8) in the entire gastrointestinal tract and in testes, compared to controls (n=8). The serum nitrite/nitrate concentration was higher in animals given LPS (n=6) than in controls (n=6).
Conclusion : After LPS, tissue clearance of albumin was not increased in any major tissue, in spite of increased serum levels of NO end products. Apparently, after activation of iNOS, the augmented release of NO is not necessarily associated with increased albumin extravasation.  相似文献   

17.
Background: Basic pharmacological research indicates that there are synergistic antinociceptive effects at the spinal cord level between adrenaline, fentanyl and bupivacaine. Our clinical experience with such a mixture in a thoracic epidural infusion after major surgery confirms this. The objectives of the present study were to evaluate the effects on postoperative pain intensity, pain relief and side effects when removing adrenaline from this triple epidural mixture. Methods: A prospective, randomised, double-blind, cross-over study was carried out in 24 patients after major thoracic or abdominal surgery. Patients with only mild pain when coughing during a titrated thoracic epidural infusion of about 10 ml · h?1 of bupivacaine 1 mg · ml?1, fentanyl 2 μg · ml?1, and adrenaline 2 μg · ml?1 were included. On the 1st and 2nd postoperative days each patient was given a double-blind epidural infusion, at the same rate, with or without adrenaline. The effect was observed for 4 h or until pain when coughing became unacceptable in spite of a rescue analgesic procedure. Rescue analgesia consisted of up to two epidural bolus injections per hour and i.v. morphine if necessary. All patients received rectal paracetamol 1 g, every 8 h. Fentanyl serum concentrations were measured with a radioimmunoassay technique at the start and end of each study period. Main outcome measures were extent of sensory blockade and pain intensity at rest and when coughing, evaluated by a visual analogue scale, a verbal categorical rating scale, the Prince Henry Hospital pain score, and an overall quality of pain relief score. Results: The number of hypaesthetic dermatomal segments decreased (P <0.001) and pain intensity at rest and when coughing increased (P <0.001) when adrenaline was omitted from the triple epidural mixture. This change started within the first hour after removing adrenaline. After 3 h pain intensity when coughing had increased to unacceptable levels in spite of rescue analgesia (epidural bolus injections and i.v. morphine). Within 15–20 min after restarting the triple epidural mixture with adrenaline, pain intensity was again reduced to mild pain when coughing. Serum concentration of fentanyl doubled from 0.22 to 0.45 ng · ml?1 (P <0.01), and there was more sedation during the period without adrenaline. Conclusions: Adrenaline increases sensory block and improves the pain-relieving effect of a mixture of bupivacaine and fentanyl infused epidurally at a thoracic level after major thoracic or abdominal surgery. Serum fentanyl concentrations doubled and sedation increased when adrenaline was removed from the epidural infusion, indicating more rapid vascular absorption and systemic effects of fentanyl.  相似文献   

18.
Blunt trauma is the principal cause of childhood death in many developed countries. This review outlines the differences between adults and children with respect to resuscitation and treatment of orthopaedic injuries in a child with polytrauma. Recent advances in techniques of fracture stabilization are reported.  相似文献   

19.
20.
Enteral feeding is often limited by gastric and intestinal motility disturbances in critically ill patients, particularly in patients with shock. So, promotility agents are frequently used to improve tolerance to enteral nutrition. This review summaries the pathophysiology, presents the available pharmacological strategies, the clinical data, the counter-indications and the principal limits. The clinical data are poor. No study demonstrates a positive effect on clinical outcomes. Metoclopramide and erythromycin seems to be the more effective. Considering the risk of antibiotic resistance, the first line use of erythromycin should be avoided in favor of metoclopramide.  相似文献   

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