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1.
目的探讨β1整合素反义寡核苷酸(ASODN)对裸鼠人胰腺癌移植瘤生长和基因表达的影响.方法建立裸鼠人胰腺癌皮下移植瘤模型,随机分为3组:对照组(皮下注射脂质体和转染液)、非特异序列(RODN)组(皮下注射RODN、脂质体和转染液)和ASODN组(皮下注射ASODN、脂质体和转染液).治疗后计算抑瘤率和肿瘤缩小率,检测肿瘤中β1整合素mRNA和蛋白表达.结果RODN组、ASODN组抑瘤率分别为4.75%和72.70%,ASODN组肿瘤缩小10.91%.与其他两组比较,ASODN组裸鼠肿瘤中β1整合素mRNA和蛋白表达水平明显降低.结论靶向β1整合素的反义寡核苷酸对人胰腺癌裸鼠皮下移植瘤的生长具有一定的抑制作用,可能为胰腺癌治疗提供新的方法.  相似文献   

2.
Wang YX  Gao L  Ji ZZ 《中华外科杂志》2005,43(21):1387-1390
目的探讨针对于胰腺癌K-ras基因点突变的反义寡脱氧核苷酸对人胰腺癌细胞株PC-2的作用。方法用脂质体将针对于K-ras基因点突变的反义寡脱氧核苷酸转染体外培养的人胰腺癌细胞株PC-2(反义组),转染正义寡脱氧核苷酸(正义组)和生理盐水(对照组),免疫组化和原位杂交检测靶基因表达,人工计数、噻唑蓝(MTT)和集落形成实验检测细胞增殖情况。结果转染48h后,反义组ras蛋白和K-ras基因mRNA的表达强度较正义组和对照组均明显降低(P〈0.05)。人工计数、MTT检测和集落实验均证实反义组细胞增殖受到明显抑制(P〈0.05)。结论针对于K-ras基因点突变的反义寡脱氧核苷酸转染体外培养的胰腺癌细胞,对靶基因表达和细胞增殖有明显的抑制作用。  相似文献   

3.
目的探讨基因枪转导突变特异性K-ras siRNA对胰腺癌细胞生长的抑制作用。方法根据胰腺癌细胞系突变特征,设计突变特异性K-ras siRNA;应用基因枪将siRNA转导入胰腺癌细胞系,提取总RNA和胞浆蛋白;应用逆转录-聚合酶链反应(RT-PCR)和免疫印迹法(Western blot)方法。检测基因枪转导突变特异性K,ras siRNA对突变型K-ras基因表达的干扰作用;行细胞爬片K-ras p21蛋白免疫组织化学染色,探讨siRNA对K-ras p21蛋白表达的抑制作用;采用CCK-8(cell countingkit,8)活细胞计数法,绘制转基因前后细胞生长曲线,观察siRNA的抑瘤效果。结果RT-PCR结果显示,K-ras突变特异性siRNA,能有效抑制K-ras基因表达(P〈0.05);Western blot和免疫组织化学结果表明,K-ras p21蛋白的表达明显降低(P〈0.05);细胞生长曲线显示,转导siRNA组较对照组细胞生长明显受抑制(P〈0.05)。结论基因枪可以有效地对小片段双链siRNA进行细胞内转导;突变特异性K-ras siRNA,可显著下调胰腺癌细胞系突变型K-ras mRNA及K-ras p21蛋白表达,并能有效抑制肿瘤细胞的增殖。  相似文献   

4.
目的 观察bcl-2硫代反义寡核苷酸(ASODN)对人恶性黑色素瘤A375细胞bcl-2mRNA及蛋白表达的影响。方法 对ASODN进行硫代修饰、脂质体转染人黑色素瘤A375细胞。实验分反义ASODN组、正义SODN组和对照组,采用逆转录聚合酶链反应(RT—PCR)和免疫组织化学检测bcl-2mRNA及蛋白水平。结果 免疫组织化学检测显示,反义组Bcl-2蛋白表达水平显著低于对照组和正义组(分别为53.14±4.26、138.22±8.45和141.08±7.83,P〈0.01);RT—PCR结果示,反义组bcl-2mRNA水平明显低于对照组和正义组(分别为0.38±0.1l、0.964-0.13和O.97±0.14,P〈0.01)。结论 bcl-2硫代反义寡核苷酸能下调人黑色素瘤A375细胞bcl-2mRNA基因水平,阻断蛋白表达。  相似文献   

5.
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。  相似文献   

6.
目的 研究生存素(survivin)反义寡核苷酸联合抑癌基因P53对人胃癌细胞系HS-746T的抑制作用及机制。方法 用P53基因和设计合成的survivin反义寡核苷酸(ASODN)对胃癌细胞系HS-746T进行处理,分空白对照组、反义survivin转染组,P53基因组,反义survivin加P53共转染组。采用细胞计数和四甲基偶氮唑蓝(MTT)法检测细胞增殖能力及细胞生长速度,用RT-PCR技术和Western印迹法分析survivin mRNA及蛋白质的表达情况,末端原位标记染色法(TUNEL)分析细胞凋亡指数。结果 不同时间反义survivin转染组、P53基因组和共转染组均对胃癌细胞的生长有抑制作用,且能够下凋胃癌细胞survivin mRNA和蛋白质的表达,共转染组较单独用药组效应明显增强,共转染组胃癌细胞凋亡指数高于另外两组。结论 Survivin反义寡核苷酸联合P53基因抑制人胃癌细胞的生长及诱导凋亡的作用大于单独应用一种药物。  相似文献   

7.
目的:探讨survivin反义寡核苷酸(ASODN)对人肝癌细胞HepG2的抑制作用。方法:采用免疫组织化学法检测肝细胞癌(HCC)组织中survivin的表达。采用脂质体介导survivin ASODN体外转染人肝癌细胞株HepG2,Western blot检测细胞survivin蛋白的表达,FCM检测细胞的凋亡率,观察细胞在软琼脂中的集落形成能力。建立人肝癌裸鼠皮下移植瘤模型,观察survivin ASODN的体内抑癌作用。 结果:(1)肝癌组织中survivin表达阳性率为75.8%(25/33),明显高于癌旁组织和正常肝组织(P<0.01);(2)survivin ASODN体外转染可明显下调HepG2细胞survivin蛋白表达,ASODN组HepG2细胞凋亡率明显高于空白对照组和SODN组(P<0.01),ASODN组HepG2细胞在软琼脂中形成的集落数目明显少于空白对照组和SODN组(P<0.01);(3)ASODN组瘤体生长速度较空白对照组和SODN组明显减慢(P<0.01),ASODN组瘤体重量较空白对照组和SODN组明显减轻(P<0.05)。结论:Survivin在HCC组织中高表达;survivin ASODN可以诱导HepG2细胞凋亡,在体外实验和动物实验中对HepG2细胞生长都有抑制作用。  相似文献   

8.
目的:比较转染Survivin、bcl -2反义寡核苷酸(ASODN)对胆囊癌细胞系 GBC SD的凋亡促进作用。方法:设计并合成特异性靶向 Survivin及 bcl- 2 的 ASODN。胆囊癌细胞分为 5 组:空白对照组、Survivin ASODN转染组、bcl- 2 ASODN转染组、Survivin SODN转染组和bcl -2 SODN转染组,转染24h后,采用逆转录 聚合酶链反应(RT PCR)检测Survivin基因、bcl- 2基因表达的变化,流式细胞仪技术检测细胞凋亡率,四甲基偶氮唑蓝(MTT)法检测 ASODN对细胞的生长抑制率。结果:转染 Survivin及bcl 2 ASODN后,胆囊癌细胞内Survivin基因表达下调 47.8%,bcl- 2 基因表达下调 52.4%;细胞凋亡率分别为(11.4±3. 9)%、(7. 3±3. 2)%,与对照组差异有统计学意义(P< 0. 01)。而转染 SurvivinASODN组及bc1 2ASODN组相比差异也有显著性(P<0.05)。两转染组相对于空白对照组的生长抑制率分别为54.3%、41.6%。结论:Survivin和bcl 2反义寡核苷酸均可有效的抑制Survivin及bc1- 2基因的表达,从而诱导胆囊癌细胞的凋亡,两者相比,Survivin反义寡核苷酸可更为有效地诱导胆囊癌细胞的凋亡。  相似文献   

9.
目的 观察核因子(NF)-кB/p65反义寡核苷酸(ASODN)对胃癌细胞增殖和凋亡的影响.方法 设计合成NF-кB/p65 ASODN,按照0.05、0.1、0.2.0.3、0.4、0.5 μmol/L不同浓度转染胃癌SGC-7901细胞株,设正义寡核苷酸(SODN)、错义寡核苷酸(MSODN)和空白对照组进行比较.利用荧光染色观察转染效果,噻唑蓝(M1T)比色法检测细胞生长抑制率(IR),流式细胞术(FCM)检测细胞凋亡情况.结果 与SODN、MSODN及对照组比较,ASODN组细胞增殖明显受到抑制(P<0.05),72 hIR为58.3%,且这种抑制作用呈浓度依赖性.FCM结果显示ASODN可诱导细胞的凋亡.结论 NF-кB ASODN能抑制胃癌细胞生长,其机制与ASODN诱导的细胞凋亡有关,NF-кB可作为胃癌治疗的新靶点.  相似文献   

10.
目的 观察Survivin ASODN联合长春新碱(VCR)、5-氟尿嘧啶(5-Fu)对人结肠癌细胞系HT-29凋亡的影响。方法 设计合成特异性Survivin反义寡核苷酸(ASODN)。HT-29细胞分成6组:空白对照组、空脂质体转染对照组、正义链转染对照组、100、200、300nmol/L反义链转染组。以阳离子脂质体为载体转染至HT-29细胞内,用Westernblot法检测各组细胞Survivin蛋白表达情况;流式细胞仪检测细胞凋亡情况;MTT法检测VCR、5-Fu对转染前后细胞的生长抑制情况。结果各浓度ASODN转染组癌细胞Survivin蛋白表达有不同程度减少,而各对照组细胞Survivin蛋白表达无明显变化。各ASODN转染组VCR、5-Fu对肿瘤细胞生长抑制率明显高于各对照组(P〈0.05),且300nmol/L转染组明显高于100和200nmol/L组(P〈0.05)。ASODN转染组细胞凋亡指数明显高于各对照组(P〈0.05),以300nmol/L转染组加VCR作用后凋亡指数最高(P〈0.01)。结论Survivin ASODN转染人结肠癌细胞能下调Survivin蛋白的表达,诱导结肠癌细胞凋亡,抑制细胞增值,增加结肠癌细胞对化疗药物的敏感性。  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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