血小板活化与兔VX2移植瘤相关性的研究
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摘要
目的：对兔VX2肝移植瘤不同时期外周血中血小板活化标志物CD62P表达水平进行监测、比较,观察血小板活化标志物CD62P表达水平的变化以及血小板活化与血栓形成、肝癌的发生、发展、浸润、转移之间的关系,探讨肿瘤细胞与血栓形成、血小板活化之间的相关性,及血小板活化在肝癌病程进展、血行转移中的重要作用,为临床肝癌的诊断、治疗、病情评估及预后等方面提供更多的理论依据,从血液学角度寻求抗癌治疗新靶点,进一步提高肝癌患者的临床诊治水平,延长肝癌患者的寿命,提高肝癌患者的生活质量。方法：1.将健康新西兰大白兔共90只,随机分为：①肝移植瘤模型组30只；②假手术组30只；③正常对照组30只；2.兔VX2肝移植瘤瘤株的复苏：从负氮瓶中取出兔VX2瘤株(瘤株由第四军医大学唐都医院动物实验中心提供),置于恒温箱中水浴复苏,复苏温度约为25-28℃,5分钟后取出复苏的瘤株,置于无菌玻璃器皿中,加入3ml含2万u庆大霉素的生理盐水,在无菌箱中将瘤株剪成1-2mm3大小的碎块,混匀制成混悬液备用；3、荷瘤兔的制备：选健康新西兰大白兔2只,固定于动物台上,选右侧臀部作为注射点,脱毛、常规消毒注射点皮肤后,用18G针头注入1ml备用的瘤细胞混悬液,拔针后压迫5分钟,防止瘤细胞混悬液从穿刺道溢出,保证瘤株种植成功。荷瘤兔种植1周后,观察荷瘤兔存活,触摸其右侧臀部已长出直径约2cm大小的包块,荷瘤兔制备成功；4、瘤细胞混悬液的制备：准备切开荷瘤兔臀部肿瘤组织,制备瘤细胞混悬液,进行传代转种。用速眠星II号将荷瘤兔全麻,无菌条件下剥离肿瘤,切开瘤体,取靠近包膜的灰白色鱼肉样组织置于生理盐水(含庆大霉素2万U)中,剔除坏死组织及纤维组织,将瘤组织剪碎成1-2mm3小块,加入生理盐水适量制成浓度约为5×1010个/L的混悬液备用。5、兔VX2肝移植瘤模型建立选健康新西兰大白兔30只由背部脊柱旁皮下注射速眠星Ⅱ号1ml,观察实验动物麻醉成功。将其仰卧位固定于动物台上,取上腹部剑突下正中线处为切口选择处,局部脱毛、常规消毒后切开皮肤,切口长约3cm左右,逐层分离皮下组织,打开腹腔,暴露肝脏。轻柔将肝脏拉出,用18G针头在肝脏左叶注入瘤细胞混悬液1ml,进针深度约2cm左右。拔针后用明胶海绵填塞、压迫穿刺道5分钟,回纳肝脏,逐层关腹。依次成功种植兔VX2肝移植瘤模型30只；6、假手术组的建立选健康新西兰大白兔30用上述同样方法麻醉后,仰卧位固定于动物台上,取上腹部剑突下正中线处为切口处,局部脱毛、常规消毒后切开皮肤,切口长约3cm左右,逐层分离皮下组织,打开腹腔,暴露肝脏。轻柔将肝脏拉出,在肝脏左叶用18G针头注入生理盐水1ml,进针深度约2cm左右。拔针后用明胶海绵填塞、压迫穿刺道5分钟,回纳肝脏,逐层关腹,依次成功操作假手组兔子30只；7、正常对照组30只健康新西兰大白兔未进行任何干预作为对照；8、采用流式细胞术(FCM)分别于瘤株种植前、瘤株种植后6h、48h、1W、2w采各组实验动物外周血,检测血小板活化标志物CD62P表达水平,比较兔肝移植瘤模型组不同时间外周血血小板活化标志物CD62P表达水平的变化,观察各组实验动物血液循环中CD62P表达水平的差异,分析转移组与无转移组CD62P表达水平的不同,所得数据用均数±标准差(x±s)表示,两样本均数比较采用成组设计的t检验,以P＜0.05为差异有统计学意义。9、在瘤株成功种植后1w、2w各组分别处死并解剖15只实验动物,肉眼观察肝脏、肺内有无转移,观察静脉内有无血栓形成；检测转移组与无转移组外周血血小板活化标志CD62P的表达水平,比较转移组及无转移组间血小板活化标志物表达水平的不同；分析有血栓形成组与无血栓形成组外周血小板活化标志物表达有无差异。结果：1、瘤株种植前各组外周血血小板活化标志物呈低表达状态,各组均数对比P值分别为0.083△0.10▲0.088°,组间对比无明显差异；2、瘤株种植后6h假手术组、肝移植瘤模型组外周血血小板活化标志物CD62P表达水平增加,假手术组与正常对照组比较P=0.01,肝移植瘤模型组与正常对照组比较P=0.012,差异均有统计学意义；肝移植瘤组与假手术组对比P=0.084,组间对比无明显变化；3、瘤株种植后48h,假手术组与正常对照组比较P=0.023,肝移植瘤组与正常对照组对比P=0.01,肝移植瘤组、假手术组血小板活化标志物的表达水平高于正常对照组。肝移植瘤组与假手术组对比P=0.013差异有统计学意义,肝移植瘤组血小板活化标志物CD62P表达高于假手术组；4、瘤株种植后1W、2w时假手术组与正常对照组比较P值均大于0.05,组间对比无统计学意义；肝移植瘤组与正常对照组、假手术组对比P值均小于0.05,兔肝移植瘤模型组外周血血小板活化标志物水平明显高于正常对照组及假手术组；5、肝移植瘤组外周血血小板活化标志物CD62P表达水平明显高于正常对照组及假手术组,曲线图示：瘤株种植后外周血小板活化标志物表达水平随着病程进展逐渐上升；正常组与假手术组曲线,在不同的检测时间外周血血小板活化标志物表达水平变化不大；假手术组曲线在种植后6h、48h稍有上升,1w后逐渐恢复,考虑与假手术组血小板活化标志物表达的短暂增加与手术创伤引有关,但其增加的幅度不大；6、瘤株种植后1W、2w后肿瘤均有转移,而且2w后检测外周血CD62P表达水平明显高于1w后,转移组与无转移组对比P值均小于0.05,差异有统计学意义,转移组的血小板活化标志物表达明显高于无转移组；7、瘤株种植后1w、2w,肝移植瘤组血小板活化标志物检测数据对比,肝内转移组与肺内转移组,肝内转移组与肝内、肺内均转移组及肺内转移组与肝内、肺内均转移组比值P值均大于0.05,组间数据对比变化不大；8、瘤株种植后1W、2w处死各组实验动物15只,发现1w后肝移植瘤组静脉血栓形成1例,2w后肝移植瘤组静脉血栓形成8例,血栓形成率约为30%；1w后假手术组静脉血栓形成1例,2w后未见血栓形成,考虑假手术组1例血栓形成与手术创伤所致凝血功能改变有关；正常对照组未见血栓形成。结论：1、兔VX2肝移植瘤瘤细胞与血小板相互作用,使血液循环中的血小板处于激活状态,其活化标志物表达水平增加；2、肿瘤的发生、发展、转移伴随着血小板活化,肿瘤中晚期外周血血小板活化标志物表达水平较早期明显增加,活化的血小板可能参与了肿瘤的浸润、转移；3、肿瘤细胞和活化的血小板相互作用,有助于血栓的形成,恶性肿瘤多伴有血栓形成；4、肿瘤细胞、血小板活化、血栓形成之间相互作用,在肝癌的发生、发展、转移中起着十分重要的作用,血小板活化标志物CD62P表达水平的检测可以作为肝癌病情进展及预后评估的指标之一。
[Abstrac] Objective:VX2 transplantation of rabbit in different periods in melanoma peripheral blood platelet activate marker, this paper discusses the monitoring, compared with liver cancer platelet activate the close relationship between the platelets and activation in liver cancer occurrence, development, and the important role of infiltration, transfer of hepatocellular carcinoma. For clinical diagnosis, treatment, illness evaluation, etc, providing new theoretical basis from blood clotting gets more treatment, and to further improve the level of diagnosis and treatment of HCC patients. For clinical liver cancer diagnosis, treatment, illness evaluation, etc, providing new theoretical basis from blood clotting gets more treatment, and to further improve the level of diagnosis and treatment of HCC patients. For clinical liver cancer diagnosis, treatment, illness evaluation, etc, providing new theoretical basis from blood clotting gets more treatment, and to further improve the level of diagnosis and treatment of HCC patients.
Methods:1. Take New Zealand big white rabbit, only 60 randomly divided into (1) liver model group; (2) JiaShouShu group; (3) normal control group.2. Tumor block anesthesia preparation:will a tumor-burdened rabbit, sterile conditions stripping tumor, cut, get close to capsule tumors RouYang organization in gray fish saline (including gentamycin 2 million U), eliminate necrotic tissue and fiber organization, will tumor tissue shear ground into 1-2mm3 small pieces. Set aside.3. tumor cells levitation liquid preparation:tumor block+ physiological saline made suspension liquid 1ml about 5x, the concentration of 1010 a/L.4, planting rabbit VX2 transplant tumor model sigmoidectomy puncture inoculation method adopted left lobe, will pull in vitro liver needle injection, with 18G (spare tumor 3-4 organization pieces) or tumor cells levitation liquid 1ml spear pierced depth about 2cm, pull needle with 1cm size gelatin sponge after 2 minutes, will block oppression liver back on, layered shut abdomen, successful planting liver rabbit model 30 only; JiaShouShu group take sigmoidectomy shap, will direct the liver pulled outside, the left lobe with 18G needles into physiological saline, 1ml pinpoint Pierce depth about 2cm, pull needle with gelatin sponge block after 2 minutes, oppression, layered on liver back will be closed abdominal (n= 30); Normal controls normal and healthy New Zealand rabbits did not undertake any intervention large (n= 30).5.adopt flow cytometric profilometry (FCM), respectively on 6 h after planting,48 h, the comprehensive in 2w 1w leds, blood, monitoring the platelet activate markers CD62p expression level, compared with standard data mean±(x±s) says, compares the two sample mean t test, the group design with P< 0.05 for difference have statistical significance. After planting in expl 1w leds,2w death 15 rabbit liver, lungs anatomic observation without metastases, of femoral vein, portal without thrombosis within. Results:1, the research team peripheral blood platelet activation markers CD62p level obviously higher than those in the control group, and the transfer of tumor progression, platelet activate marker has risen,2 weeks of peak; 2.after planting anatomical rabbit, delayed-type 1w leds not seen transfer and thrombosis. Double lung and after 2w team are JiaShouShu liver metastases,2 cases of thrombosis, the normal group had not changed. Research group and control group, and compared using t-test between groups, (t= 0.18) (P < 0.01) difference was statistically significant. Conclusion:VX2 rabbit transplant tumor peripheral blood platelet is active with tumors, and the occurrence, development and transfer of platelet activate markers expression was increased. Platelet activate markers detection can be used as tumor progression, transfer effective evaluation index.
[Abstrac] Objective:VX2 transplantation of rabbit in different periods in melanoma peripheral blood platelet activate marker, this paper discusses the monitoring, compared with liver cancer platelet activate the close relationship between the platelets and activation in liver cancer occurrence, development, and the important role of infiltration, transfer of hepatocellular carcinoma. For clinical diagnosis, treatment, illness evaluation, etc, providing new theoretical basis from blood clotting gets more treatment, and to further improve the level of diagnosis and treatment of HCC patients. For clinical liver cancer diagnosis, treatment, illness evaluation, etc, providing new theoretical basis from blood clotting gets more treatment, and to further improve the level of diagnosis and treatment of HCC patients. For clinical liver cancer diagnosis, treatment, illness evaluation, etc, providing new theoretical basis from blood clotting gets more treatment, and to further improve the level of diagnosis and treatment of HCC patients.
Methods:1. Take New Zealand big white rabbit, only 60 randomly divided into (1) liver model group; (2) JiaShouShu group; (3) normal control group.2. Tumor block anesthesia preparation:will a tumor-burdened rabbit, sterile conditions stripping tumor, cut, get close to capsule tumors RouYang organization in gray fish saline (including gentamycin 2 million U), eliminate necrotic tissue and fiber organization, will tumor tissue shear ground into 1-2mm3 small pieces. Set aside.3. tumor cells levitation liquid preparation:tumor block+ physiological saline made suspension liquid 1ml about 5x, the concentration of 1010 a/L.4, planting rabbit VX2 transplant tumor model sigmoidectomy puncture inoculation method adopted left lobe, will pull in vitro liver needle injection, with 18G (spare tumor 3-4 organization pieces) or tumor cells levitation liquid 1ml spear pierced depth about 2cm, pull needle with 1cm size gelatin sponge after 2 minutes, will block oppression liver back on, layered shut abdomen, successful planting liver rabbit model 30 only; JiaShouShu group take sigmoidectomy shap, will direct the liver pulled outside, the left lobe with 18G needles into physiological saline, 1ml pinpoint Pierce depth about 2cm, pull needle with gelatin sponge block after 2 minutes, oppression, layered on liver back will be closed abdominal (n= 30); Normal controls normal and healthy New Zealand rabbits did not undertake any intervention large (n= 30).5.adopt flow cytometric profilometry (FCM), respectively on 6 h after planting,48 h, the comprehensive in 2w 1w leds, blood, monitoring the platelet activate markers CD62p expression level, compared with standard data mean±(x±s) says, compares the two sample mean t test, the group design with P< 0.05 for difference have statistical significance. After planting in expl 1w leds,2w death 15 rabbit liver, lungs anatomic observation without metastases, of femoral vein, portal without thrombosis within. Results:1, the research team peripheral blood platelet activation markers CD62p level obviously higher than those in the control group, and the transfer of tumor progression, platelet activate marker has risen,2 weeks of peak; 2.after planting anatomical rabbit, delayed-type 1w leds not seen transfer and thrombosis. Double lung and after 2w team are JiaShouShu liver metastases,2 cases of thrombosis, the normal group had not changed. Research group and control group, and compared using t-test between groups, (t= 0.18) (P < 0.01) difference was statistically significant. Conclusion:VX2 rabbit transplant tumor peripheral blood platelet is active with tumors, and the occurrence, development and transfer of platelet activate markers expression was increased. Platelet activate markers detection can be used as tumor progression, transfer effective evaluation index.
引文
1.丁水康,肝癌的流行因素和预防[J],Journal of clinical and Experimental medicine.2009,8(2):118.
2.高蒿,杨仁杰等.肝癌综合介入治疗的临床研究进展[J],临床医药时间杂志.2008,17(9)：721-723.
3.孙璐,曹贵文等.TACE联合I125放射性粒子植入术治疗肝癌临床研究[J].潍坊医学学报.2008.30(5)：445.
4.王杰军,肿瘤转移机制及诊断进展[J].上海：第二军医大学出版社,2002：270-272.
5. Jurk k,kehrel BE.Platelets:physiology and biochemistry [J]. Semin Thromb Hemost,2005,31:381-392.
6. Michelson AD,Bamaard MR. Evaluation of platelet function by flow cytomet ry[J]. Methods,2000,21 (3):259-270.
7.张峻梅,唐方,李玲.恶性肿瘤患者活化血小板的检测[J].中国肿瘤临床,2005,32(16)：920-922.
8.向琪,刘敏娟.肺癌和卵巢癌患者血栓相关标志物的实验研究[J].血栓与止血学,2010,16(1)：27-29.
9.练练,陶敏,张有涛,等.肺癌患者外周血小板活化指标测定及临床意义[J].中国血液流变学杂志,2010；20(1)：114-116.
10. Jurk k,kehrel BE.Platelets:physiology and biochemistry [J]. Semin Thromb Hemost,2005,31:381-392.
11.郭金英,陈彦,文阳安.等.肿瘤患者外周血活化血小板测定机其临 出意义[J].重庆医科大学学报,2008,33(4)：510-512.
12.张爱华,李子亮,孙达春,等.介入治疗前后凝血及纤溶功能变化及临床意义[J].复旦学报：医学版,2005.32(1)18-20.
13. sun DC,zhang AH.Li xl,et al.Effect of platelet actvation and endothelial cell injury on malignant cancer [J].The Chinese-German J Clin oncol,2003,2(4):243-245.
14. Leo, Pratico D,LUliano L,et al.Platelet activation by superoxide anion and hydroxyl radicals intrinsicals intrsically generated by platelets that had undergone anoxia and t han reoxygenated[J] circulatin, 1997,95(4):885-891.
15.赵益明.活化血小板分子标志物：P选择素、糖盏蛋白和糖蛋白V的研究进展[J].血栓与止血学,2005,11(2)：85-87.
16.杨树君,战芹等.卵巢癌患者外周血小板活化分子CD62P, CD63P含量的临床检测及意义[J].中国实验诊断学.2001.5(5)：256.
17.陈艳,王熙才,伍治平,等.肝癌患者外周血血小板活化标志物检测及临床意义[J].中华肿瘤防治杂志,2006,13(18)：1412-1415.
18. Gupta GP,Massague J.Platelets and metastasis revisited:a novel fatty link [J] J Clin Invest,2004,114:1691-1693.
19.赵益明。活化血小板分子标志物：P-选择素、唐盏蛋白和糖蛋白v的研究进展[J].血栓与止血学,2005,11(2)：85-87.
20.P.赫尔曼尼克,杨勇翻译.国际抗癌联盟肿瘤TNM分期图谱[M].北京：科学出版社,2002：81-92.
21.刘元波,赵相印。肿瘤所致凝血异常的研究进展[J].首都医科大学学报,2000,21(3)：270-272.
22.王学锋,王鸿利.恶性肿瘤与血栓形成[J].J Diagn concepts pract, 2002,2:112-115.
23.张斌,胡毅敏,丁连举等.恶性肿瘤患者血小板检测及临床评价[J].中国民族民间医药,2010,4(1)：88.
24. Ferroni P,Roselli M,Martini F.et al.Prognostic Value of soluble P-selectin levels in colorectal cancer [J].In t cancer,2004,11(3): 404-408.
25.伍治平,王熙才,金从国,等。消化道肿瘤患者外周血血小板活化标志物检测及临床意义[J].肿瘤研究与临床,2005,17(2)：101-103.
26. Yu Y,Zhou x D,liu Y K,et al.Platelets promote the adhesion of human hepatoms cells with a highly metastatic protential to extracellular matrix protion:Involvement of platelet P-selectin and GP II b-IIIa[J].J Cancer Res Clin Oncol,2002,128(5):283-287.
27.何斌,顾健,石建霞,等.VEGF, CD62P和PMA与SLE患者易栓状态关系研究[J].血栓与止血学.2008,14(4)：157-159.
28.邹丽芳,胡钧培.恶性肿瘤与止凝血功能[J].血栓与止血学.2008,14(2)：83-84.
29. Gupta GP,Massague J.Platelets and metastasis revisited:anvovel fatty link [J].J CLin Invest,2004,114(12):1691-1693.
30. Dardik R,Savion N,Kaufman Y,et al.Thrombin promotes platelet-mediated mediated melanoma cell adhesion to endothelial cells under flow conditions:role of platelet glycoproteins P-Selectin and GP Ⅱ b-Ⅲ A[J].Br J Cancer,1998,77(12):2069-2075.
31.冯志平,李积德,李进章.血小板与肿瘤的关系[J].牡丹江医学学报,2007,28(6)72-74.
1.丁水康,肝癌的流行因素和预防[J] Journal of clinical and Experimental medicine.2009;8(2);118.
2.高 嵩综述,杨仁杰等肝癌综合介入治疗的临床研究进展[J],临床医药实践杂志.2008；17(9)；721-723.
3.王建华,肝癌综合介入治疗的现状[J]中华肝脏病杂志2005,13(10)；721-723.
4.孙璐,曹贵文等.TACE联合125I放射性粒子植入术治疗肝癌临床研究[J].潍坊医学学报。2008.30(5)；445
5.胡洋.肝动脉联合门静脉栓塞化疗治疗巨块型肝癌的临床分析.HEILONGJIAN MEDICINE AND PHARMACY.2009.32(2);53
6.吴沛宏等,肝癌微创治疗与多学科综合治疗,2003,10(1)
7. Rossi S,Buscarini E,Garbagnati F,et al.percutaneous treatmentof small hepatic tumors by an expandable RF needle electrode [J].Am J Roentgenol,1998,170(10); 1015.
8.黎海亮,胡鸿涛.肝癌非手术治疗进展河南医学研究2008,17(3)：266-269
9.徐向东,朱裕辉等,微波治疗肝癌的临床研究[J],中外医疗,2008,35：38-39
10.吴沛宏、张福君等.肝癌微创治疗与多学科综合治疗.北京：军事医学科学出版社,2003：183-187.
11.Pacella C M, Bizzarri G, Cecconi P, et al. Hepatoeellular carcinoma longterm results of combined treatment with laser thermal ablatio and transcatheterterial chemoembolization[J]. Radiology, 2001,219(3)669-679.
12.郑家平,肝癌局部热毁治疗进展[J],Cllinese J Of rOday'S Med.2003, 3(5):112-115
13.钱林学,魏红涛,胡向东肝癌局部消融治疗的现状和进展世界华人消化杂志2008；16(18)：1955—1961
14.张金山,TACE联合其他手段治疗肝癌发展现状[J]肿瘤进展学.2006,13(2)：856-857
15.程树群,周信达,汤钊猷,等.磺化油与高功能聚焦超声破坏肝组织的协同升温效应研究[J].中国超吉医学杂志,1997,13(4)：1—4.
16.赵建农,王智彪,郭大静,等.肝癌经肝动脉碘油化疗药物拴塞联合高强度聚焦超声治疗后的CT评价及临床意义[J].中华肝脏病杂志,2001,7(9)：61-63.
17. Koda M, Murawak Y, M itsuda A, et al. Combinationtherapy with transcatheter arterial chemoem—bolization and percutaneous ethanol injection alonbolization and percutaneous ethanol injection aloncarcinoma:a randomized control study[J]. Cancer,2001,92(6): 1516-1524
18. ChiaHsien C J, Chuang V P。Cheng S H, et al. Unresectable hepatocellular carcinoma treated with radiotherapy and chemoembolization[J]. Int J Cancer2001,96(4):243-252
19.沈建东,化疗栓塞联合三维适行放疗治疗原发性肝癌[J],医学信息手术学分册；2008,21(11)：1034-1036
20.李进贤,薛涣洲,陈汝福,等.原发性肝癌介入治疗前后血清血管内皮生长因子的变化[J].中华实验外科杂志,2002,19(5)：417—418.
21. Kim Y B, Park Y N, Park C, et al. Increased proliferation activaties of vascular enmour cells in residual heDatocel] ular carcinoma foldothelial cells and tulowing transcatheter arterial embolization [j]. HistopatholOgy,2001,38(2):160-166.
22.刘秀芳,张海,王炳胜,等.沙利度胺联合介入治疗中晚期肝癌疗效观察[J].中国肿瘤临床与康复,2007。14(6)：546—550.
23.于志坚,孟宪墉.HCPT, CDDP,5-FU等肝动脉栓塞灌注治疗原发性肝癌86例[J].中国肿瘤临床,1998,25(1)：73—75.
24.郝强,田建明,曹雪涛,等.经肝动脉插管注射重组腺病毒治疗大鼠转移性肝癌的实验研究[J].中华放射学杂志,2000,34(10)：666669.
25.吴沛宏,黄金华,罗鹏飞,等.肿瘤介入诊疗学[M].北京：人民军医出版,2005：625—664.
26.花勤亮,崔艳慧,李四清,等.肝动脉化疗栓塞结合三维适形放疗治疗原发性肝癌[J].现代肿瘤医学,2006,14(10)：1266—1269.
27. Llovet J, Riei S。Mazzaferro V, et al. Sorafenib improves survival inadvanced hepatocellular carcinoma(HCC):results of a Phase III randomized placebo—controlled trial (SI Ptrial), Proceedingsfromthe AmericanSocietyofCIiniealOn—cology Conference. Chicago. IL. 2007,13(2):2221-2223.
28.夏锋,王曙光,等。肝癌治疗的新观念[J].实用临床医药杂志,2009,13 (2) ：1-3.
29.彭磷基,陆大祥。肝癌治疗学的研究进展[J]。中国老年学杂志,2009,3(29)：628-630。
2.高蒿,杨仁杰等.肝癌综合介入治疗的临床研究进展[J],临床医药时间杂志.2008,17(9)：721-723.
3.孙璐,曹贵文等.TACE联合I125放射性粒子植入术治疗肝癌临床研究[J].潍坊医学学报.2008.30(5)：445.
4.王杰军,肿瘤转移机制及诊断进展[J].上海：第二军医大学出版社,2002：270-272.
5. Jurk k,kehrel BE.Platelets:physiology and biochemistry [J]. Semin Thromb Hemost,2005,31:381-392.
6. Michelson AD,Bamaard MR. Evaluation of platelet function by flow cytomet ry[J]. Methods,2000,21 (3):259-270.
7.张峻梅,唐方,李玲.恶性肿瘤患者活化血小板的检测[J].中国肿瘤临床,2005,32(16)：920-922.
8.向琪,刘敏娟.肺癌和卵巢癌患者血栓相关标志物的实验研究[J].血栓与止血学,2010,16(1)：27-29.
9.练练,陶敏,张有涛,等.肺癌患者外周血小板活化指标测定及临床意义[J].中国血液流变学杂志,2010；20(1)：114-116.
10. Jurk k,kehrel BE.Platelets:physiology and biochemistry [J]. Semin Thromb Hemost,2005,31:381-392.
11.郭金英,陈彦,文阳安.等.肿瘤患者外周血活化血小板测定机其临 出意义[J].重庆医科大学学报,2008,33(4)：510-512.
12.张爱华,李子亮,孙达春,等.介入治疗前后凝血及纤溶功能变化及临床意义[J].复旦学报：医学版,2005.32(1)18-20.
13. sun DC,zhang AH.Li xl,et al.Effect of platelet actvation and endothelial cell injury on malignant cancer [J].The Chinese-German J Clin oncol,2003,2(4):243-245.
14. Leo, Pratico D,LUliano L,et al.Platelet activation by superoxide anion and hydroxyl radicals intrinsicals intrsically generated by platelets that had undergone anoxia and t han reoxygenated[J] circulatin, 1997,95(4):885-891.
15.赵益明.活化血小板分子标志物：P选择素、糖盏蛋白和糖蛋白V的研究进展[J].血栓与止血学,2005,11(2)：85-87.
16.杨树君,战芹等.卵巢癌患者外周血小板活化分子CD62P, CD63P含量的临床检测及意义[J].中国实验诊断学.2001.5(5)：256.
17.陈艳,王熙才,伍治平,等.肝癌患者外周血血小板活化标志物检测及临床意义[J].中华肿瘤防治杂志,2006,13(18)：1412-1415.
18. Gupta GP,Massague J.Platelets and metastasis revisited:a novel fatty link [J] J Clin Invest,2004,114:1691-1693.
19.赵益明。活化血小板分子标志物：P-选择素、唐盏蛋白和糖蛋白v的研究进展[J].血栓与止血学,2005,11(2)：85-87.
20.P.赫尔曼尼克,杨勇翻译.国际抗癌联盟肿瘤TNM分期图谱[M].北京：科学出版社,2002：81-92.
21.刘元波,赵相印。肿瘤所致凝血异常的研究进展[J].首都医科大学学报,2000,21(3)：270-272.
22.王学锋,王鸿利.恶性肿瘤与血栓形成[J].J Diagn concepts pract, 2002,2:112-115.
23.张斌,胡毅敏,丁连举等.恶性肿瘤患者血小板检测及临床评价[J].中国民族民间医药,2010,4(1)：88.
24. Ferroni P,Roselli M,Martini F.et al.Prognostic Value of soluble P-selectin levels in colorectal cancer [J].In t cancer,2004,11(3): 404-408.
25.伍治平,王熙才,金从国,等。消化道肿瘤患者外周血血小板活化标志物检测及临床意义[J].肿瘤研究与临床,2005,17(2)：101-103.
26. Yu Y,Zhou x D,liu Y K,et al.Platelets promote the adhesion of human hepatoms cells with a highly metastatic protential to extracellular matrix protion:Involvement of platelet P-selectin and GP II b-IIIa[J].J Cancer Res Clin Oncol,2002,128(5):283-287.
27.何斌,顾健,石建霞,等.VEGF, CD62P和PMA与SLE患者易栓状态关系研究[J].血栓与止血学.2008,14(4)：157-159.
28.邹丽芳,胡钧培.恶性肿瘤与止凝血功能[J].血栓与止血学.2008,14(2)：83-84.
29. Gupta GP,Massague J.Platelets and metastasis revisited:anvovel fatty link [J].J CLin Invest,2004,114(12):1691-1693.
30. Dardik R,Savion N,Kaufman Y,et al.Thrombin promotes platelet-mediated mediated melanoma cell adhesion to endothelial cells under flow conditions:role of platelet glycoproteins P-Selectin and GP Ⅱ b-Ⅲ A[J].Br J Cancer,1998,77(12):2069-2075.
31.冯志平,李积德,李进章.血小板与肿瘤的关系[J].牡丹江医学学报,2007,28(6)72-74.
1.丁水康,肝癌的流行因素和预防[J] Journal of clinical and Experimental medicine.2009;8(2);118.
2.高 嵩综述,杨仁杰等肝癌综合介入治疗的临床研究进展[J],临床医药实践杂志.2008；17(9)；721-723.
3.王建华,肝癌综合介入治疗的现状[J]中华肝脏病杂志2005,13(10)；721-723.
4.孙璐,曹贵文等.TACE联合125I放射性粒子植入术治疗肝癌临床研究[J].潍坊医学学报。2008.30(5)；445
5.胡洋.肝动脉联合门静脉栓塞化疗治疗巨块型肝癌的临床分析.HEILONGJIAN MEDICINE AND PHARMACY.2009.32(2);53
6.吴沛宏等,肝癌微创治疗与多学科综合治疗,2003,10(1)
7. Rossi S,Buscarini E,Garbagnati F,et al.percutaneous treatmentof small hepatic tumors by an expandable RF needle electrode [J].Am J Roentgenol,1998,170(10); 1015.
8.黎海亮,胡鸿涛.肝癌非手术治疗进展河南医学研究2008,17(3)：266-269
9.徐向东,朱裕辉等,微波治疗肝癌的临床研究[J],中外医疗,2008,35：38-39
10.吴沛宏、张福君等.肝癌微创治疗与多学科综合治疗.北京：军事医学科学出版社,2003：183-187.
11.Pacella C M, Bizzarri G, Cecconi P, et al. Hepatoeellular carcinoma longterm results of combined treatment with laser thermal ablatio and transcatheterterial chemoembolization[J]. Radiology, 2001,219(3)669-679.
12.郑家平,肝癌局部热毁治疗进展[J],Cllinese J Of rOday'S Med.2003, 3(5):112-115
13.钱林学,魏红涛,胡向东肝癌局部消融治疗的现状和进展世界华人消化杂志2008；16(18)：1955—1961
14.张金山,TACE联合其他手段治疗肝癌发展现状[J]肿瘤进展学.2006,13(2)：856-857
15.程树群,周信达,汤钊猷,等.磺化油与高功能聚焦超声破坏肝组织的协同升温效应研究[J].中国超吉医学杂志,1997,13(4)：1—4.
16.赵建农,王智彪,郭大静,等.肝癌经肝动脉碘油化疗药物拴塞联合高强度聚焦超声治疗后的CT评价及临床意义[J].中华肝脏病杂志,2001,7(9)：61-63.
17. Koda M, Murawak Y, M itsuda A, et al. Combinationtherapy with transcatheter arterial chemoem—bolization and percutaneous ethanol injection alonbolization and percutaneous ethanol injection aloncarcinoma:a randomized control study[J]. Cancer,2001,92(6): 1516-1524
18. ChiaHsien C J, Chuang V P。Cheng S H, et al. Unresectable hepatocellular carcinoma treated with radiotherapy and chemoembolization[J]. Int J Cancer2001,96(4):243-252
19.沈建东,化疗栓塞联合三维适行放疗治疗原发性肝癌[J],医学信息手术学分册；2008,21(11)：1034-1036
20.李进贤,薛涣洲,陈汝福,等.原发性肝癌介入治疗前后血清血管内皮生长因子的变化[J].中华实验外科杂志,2002,19(5)：417—418.
21. Kim Y B, Park Y N, Park C, et al. Increased proliferation activaties of vascular enmour cells in residual heDatocel] ular carcinoma foldothelial cells and tulowing transcatheter arterial embolization [j]. HistopatholOgy,2001,38(2):160-166.
22.刘秀芳,张海,王炳胜,等.沙利度胺联合介入治疗中晚期肝癌疗效观察[J].中国肿瘤临床与康复,2007。14(6)：546—550.
23.于志坚,孟宪墉.HCPT, CDDP,5-FU等肝动脉栓塞灌注治疗原发性肝癌86例[J].中国肿瘤临床,1998,25(1)：73—75.
24.郝强,田建明,曹雪涛,等.经肝动脉插管注射重组腺病毒治疗大鼠转移性肝癌的实验研究[J].中华放射学杂志,2000,34(10)：666669.
25.吴沛宏,黄金华,罗鹏飞,等.肿瘤介入诊疗学[M].北京：人民军医出版,2005：625—664.
26.花勤亮,崔艳慧,李四清,等.肝动脉化疗栓塞结合三维适形放疗治疗原发性肝癌[J].现代肿瘤医学,2006,14(10)：1266—1269.
27. Llovet J, Riei S。Mazzaferro V, et al. Sorafenib improves survival inadvanced hepatocellular carcinoma(HCC):results of a Phase III randomized placebo—controlled trial (SI Ptrial), Proceedingsfromthe AmericanSocietyofCIiniealOn—cology Conference. Chicago. IL. 2007,13(2):2221-2223.
28.夏锋,王曙光,等。肝癌治疗的新观念[J].实用临床医药杂志,2009,13 (2) ：1-3.
29.彭磷基,陆大祥。肝癌治疗学的研究进展[J]。中国老年学杂志,2009,3(29)：628-630。