摘要
研究背景与目的:皮肤软组织扩张术已广泛应用于临床,成为整形外科常规治疗手段之一,用于多种组织缺损的修复和器官再造。但是在扩张器植入术后经常会出现一些并发症,如血肿、感染、皮瓣坏死等,其中血肿是最常见的。血肿不仅会对皮瓣产生压迫使组织缺血,而且会对组织产生毒性作用。此外,血肿又与感染和皮瓣坏死密切相关。目前,临床上对于扩张器植入术后血肿的治疗仅限于清除血肿,尚无有效的减轻皮瓣损伤的治疗药物。本研究的目的是探讨非张力性血肿引起扩张器表面皮瓣损伤的机制,为减轻血肿造成的皮瓣损伤寻求有效的治疗药物,并探讨其作用机理。本研究由两部分组成:
第一部分去铁胺减轻非张力性血肿中铁对未扩张皮瓣毒性作用的研究
方法:13头4～8月龄白色小型猪,在每头猪脊柱两侧各设计5个术区,共128个术区,共植入128个80ml肾形扩张器,形成128个未扩张皮瓣。将未扩张皮瓣随机分配进入对照组、血肿组、去铁胺组、阿加曲班组,每组32个未扩张皮瓣。其中关于阿加曲班组的实验内容将在论文第二部分的实验二中叙述。对照组:扩张器植入后不做其他处理;血肿组:在扩张器植入后向剥离腔隙内注入15ml自体血液;去铁胺组:在扩张器植入后向剥离腔隙内注入15ml自体血液和270mg甲磺酸去铁胺。各组分别在术后24h、72h、120h取材,每组共取材24个未扩张皮瓣。每组取材后剩余的8个扩张器于术后8天注水,注水完毕后取材。检测各组扩张器植入术后2h皮瓣的血流情况,检测未扩张皮瓣铁含量、丙二醛(MDA)含量、髓过氧化物酶(MPO)活性、扩张皮瓣包膜厚度、包膜胶原含量和扩张皮瓣即时回缩率。
结果:(1)三个组扩张器植入术后2h未扩张皮瓣血流灌注量无显著差别(P＞0.05);(2)血肿组术后皮瓣铁含量、MDA含量和MPO活性均高于对照组(P＜0.01);去铁胺组皮瓣铁含量和MDA含量低于血肿组(P＜0.01),MPO活性与血肿组无显著差异(P＞0.05);(3)血肿组扩张皮瓣包膜厚度、包膜胶原含量和扩张皮瓣即时回缩率均高于对照组(P＜0.01),去铁胺组与血肿组无显著差异(P＞0.05)。
第二部分阿加曲班减轻非张力性血肿中凝血酶对扩张皮瓣的影响及其机制的研究
实验一凝血酶对猪皮肤成纤维细胞增殖作用的研究
方法:采用贴壁法进行猪皮肤成纤维细胞原代培养,并传代、鉴定。用MTT法检测不同浓度凝血酶刺激后的成纤维细胞活力。用氯胺T法检测不同浓度凝血酶刺激后的细胞上清中羟脯氨酸含量。用免疫组化的方法检测凝血酶刺激后的成纤维细胞中α-平滑肌肌动蛋白(α-SMA)的表达情况。在培养液中加入阿加曲班后重复上述检测。
结果:(1)培养的细胞为典型的成纤维细胞形态,波形蛋白单克隆抗体免疫组化染色呈阳性。(2)加入不同浓度凝血酶的各组吸光值均高于不加凝血酶组(A组)(P＜0.01),加入阿加曲班和凝血酶的各组吸光值与A组无显著性差异(P＞0.05)。(3)加入不同浓度凝血酶的各组细胞上清中羟脯氨酸含量均高于不加凝血酶组(A组)(P＜0.01),加入阿加曲班和凝血酶的各组上清中羟脯氨酸含量与A组相比无统计学差异(P＞0.05)。(4)凝血酶刺激后的成纤维细胞中有α-SMA的表达,加入阿加曲班后此种表达被抑制。
实验二阿加曲班减轻非张力性血肿中凝血酶对扩张皮瓣影响的研究
方法:本实验设计对照组、血肿组和阿加曲班组。其中对照组和血肿组的实验设计与第一部分中的对照组和血肿组相同。阿加曲班组:在扩张器植入后向剥离腔隙内注入15ml血液和500μg阿加曲班。各组分别在术后24h、72h、120h取材,每组取材后剩余的8个扩张器于术后8天注水,注水完毕后取材。检测未扩张皮瓣MPO活性、MDA含量、扩张皮瓣包膜厚度、包膜胶原含量和扩张皮瓣即时回缩率。用免疫组化的方法检测未扩张皮瓣中细胞间粘附分子-1(ICAM-1)和扩张皮瓣中α-SMA的表达情况。
结果:(1)血肿组术后皮瓣MPO活性和ICAM-1的表达量均高于对照组(P＜0.01),阿加曲班组皮瓣MPO活性和ICAM-1的表达量均低于血肿组(P＜0.01);(2)血肿组术后72h和120h皮瓣MDA含量均高于对照组(P＜0.01),阿加曲班组术后72h和120h皮瓣MDA含量低于血肿组(P＜0.05)(3)血肿组扩张皮瓣包膜厚度、包膜胶原含量、扩张皮瓣中α-SMA的表达量、扩张皮瓣即时回缩率均高于对照组(P＜0.05),阿加曲班组的这些指标均低于血肿组(P＜0.05)。
结论:
1.扩张器植入术后形成的非张力性血肿能使未扩张皮瓣脂质过氧化反应和炎症反应增强。局部应用去铁胺能减轻血肿造成的脂质过氧化反应,去铁胺的这种作用主要是通过降低皮瓣中的铁含量而实现的。局部应用阿加曲班能减轻血肿造成的脂质过氧化反应和炎症反应,阿加曲班的这种作用是通过抑制血肿中的凝血酶而实现的。
2.凝血酶能诱导成纤维细胞增殖、胶原分泌量增加,能刺激成纤维细胞表达α-SMA,使成纤维细胞转变为肌成纤维细胞。阿加曲班能抑制凝血酶的这种作用。
3.非张力性血肿能使扩张皮瓣的纤维包膜增厚、包膜胶原含量增加、α-SMA表达增多、扩张皮瓣即时回缩率增加。局部应用去铁胺不能减轻血肿的这种作用。局部应用阿加曲班能减轻血肿的这种作用,阿加曲班的这种作用是通过抑制血肿中的凝血酶而实现的。
Background and Objective:Skin soft tissue expansion,which was a conventional therapy for reparation of tissue defect and organ reconstruction in plastic surgery,had been used in clinic generally.But some complications,such as: hematoma,infection,skin flap necrosis and etc,appeared after skin soft tissue expansion.Hematoma was the most frequent complication.Hematoma could not only make skin flap ischemic,but have toxic effects on skin flap.Moreover,hematoma was related to infection and skin flap necrosis closely.At present,removing the sludged blood was the only method to treat hematoma formed after skin soft tissue expansion.There was no potent medicine to release the skin flap's impairment.The objective of our research were exploring the mechanisms which the hematoma causes the skin flap's impairment,and seeking the medicines which can lessen the effect on the skin flap's impairment and exploring the medicines' mechanism of action.
PartⅠThe study of deferoxamine lessening the toxic effects of iron in non-compressive hematoma on pre-expanding skin flap
Methods:Thirteen 4～8-month-aged white minipigs were selected.10 operation zones were designed on two side of vertebral column in each minipig.The total operation zones were 128.128 kidney-shaped expanders were implanted.128 pre-expanding skin flaps were formed.The skin flaps were divided into control group, hematoma group,deferoxamine group and argatroban group randomly.The content about argatroban group would describe in PartⅡ.The control group:there was no other treatment after expander implantation.The hematoma group:15ml blood was infused into the dissected capsules after expander implantation.The deferoxamine group:15ml blood and 270mg deferoxamine were infused into the dissection capsules after expander implantation.Seperately,pieces of skin flap tissue were taken from the pre-expanding skin flaps 24h,72h,120h after soft tissue expansion.8 days after the operation,normal sodium was injected into the 8 remained expanders.Two months later,Pieces of tissue were taken from the expanded skin flap.The blood flow of pre-expanding skin flaps were detected 2h after the operation.The contents of iron and malonaldehyde(MDA),the activity of myeloperoxidase(MPO),the thickness and collagen content of the expanded skin flap's capsule,immediate contraction rate of expanded skin flap were detected.
Results:(1) The blood flow of pre-expanding skin flaps which were detected 2h after the operation had no significant difference among the three groups(P＞0.05). (2)Compared with the control group,the contents of iron and MDA and the activity of MPO in hematoma group increased(P＜0.01).Compared with the hematoma group, the contents of iron and MDA in deferoxamine group decreased(P＜0.01).The activity of MPO had no significant difference between hematoma group and deferoxamine group(P＞0.05).(3) Compared with the control group,the thickness and collagen content of the expanded skin flap's capsule and immediate contraction rate of expanded skin flap in hematoma group increased(P＜0.01).The thickness and collagen content of the expanded skin flap's capsule and immediate contraction rate of expanded had no significant difference between hematoma group and deferoxamine group(P＞0.05).
PartⅡStudy of argatroban lessening the effects of thrombin in noncompressive hematoma on expanded skin flap and the mechanisms
ExperimentⅠStudy of thrombin on proliferation of fibroblasts isolated from mini pig's skin
Methods:The minipig skin fibroblasts were cultured in monolayer.The vigor of fibroblasts which were stimulated by different concentration of thrombin was detected by MTT.The contents of hydroxyproline in supernatant were detected by chloramines T method.The expressions ofα-SMA in fibroblasts which were stimulated by thrombin,were detected by immunohistochemistry staining.After argatroban was infused into the culture solution,the detections mentioned above were repeated.
Results:(1) The cells appeared to be the typical fibroblast,the immunohistochemistry staining ofα-SMA in the cells were positive.(2) Compared with the group without thrombin,the OD value of the groups contained different concentration of thrombin were increased(P＜0.01),the OD value of the groups contained different concentration of thrombin and argatroban had no changes(P＞0.05).(3) Compared with the group without thrombin,the hydroxyproline content in supematant of the groups contained different concentration of thrombin were increased(P＜0.01),the hydroxyproline content in supernatant of the groups contained different concentration of thrombin and argatroban had no changes(P＞0.05).(4)α-SMA expressed in the fibroblast stimulated by thrombin.The expression ofα-SMA was inhibited by argatroban.
ExperimentⅡStudy of argatroban lessening the effects of thrombin in noncompressive hematoma on expanded skin flap
Methods:This experiment had three groups:control group,hematoma group, argatroban group.The dsigns of control group and hematoma group were same to the first part.The argatroban group:15ml and 500μg argatroban blood were infused into dissected capsule after expander implantation.Seperately,pieces of skin flap tissue were taken from the pre-expanding skin flaps 24h,72h,120h after soft tissue expansion.8 days after the operation,normal sodium was injected into the 8 remained expanders.Two months later,Pieces of tissue were taken from the expanded skin flap. The contents of MDA,the activity of MPO,the thickness and collagen content of the expanded skin flap's capsule,immediate contraction rate of expanded skin flap were detected.The expression of ICAM-1 in pre-expanding skin flaps andα-SMA in expanded skin flaps were detected by immunohistochemistry staining.
Results:(1)Compared with control group,the activy of MPO and expression of ICAM-1 in hematoma group increased(P＜0.01).Compared with hematoma group, the activy of MPO and expression of ICAM-1 in argatroban group decreased(P＜0.01).(2) Compared with control group,the content of MDA in hematoma group 72h and 120h after operation increased(P＜0.01).Compared with hematoma group,the content of MDA in argatroban group 72h and 120h after operation decreased(P＜0.01).(3) Compared with control group,the thickness and collagen content of the expanded skin flap's capsule,immediate contraction rate of expanded skin flap,the expression ofα-SMA in hematoma group increased(P＜0.01).Compared with hematoma group,these indexes in argatroban group decreased(P＜0.01).
Conclusions
1.The hematoma which formed after skin soft tissue expansion can strengthen the lipid peroxidation and inflammatory reaction in the skin flap.Using deferoxamine locally can reduce the lipid peroxidation caused by hematoma.This role of deferoxamine results from decreasing the content of iron in the skin flap.Using argatroban locally can reduce the lipid peroxidation and inflammatory reaction caused by hematoma.This role of argatroban results from inhibiting thrombin in hematoma.
2.Thrombin can induce fibroblast's proliferation,make fibroblast product more collagen,increase the expression ofα-SMA in fibroblast,transform fibroblast to myofibroblast.Argatroban can inhibit these effects of thrombin.
3.Hematoma can make the capsular thickness thicken,increase collagen content of the capsule and immediate contraction rate of expanded skin flap.Using deferoxamine locally can not lessen these effects of hematoma.Using argatroban locally can lessen these effects of hematoma.This role of argatroban is result from inhibiting thrombin in hematoma.
引文
1.Angel MF,Haddad J,Abramson M.A free radical scavenger reduced hematoma-induced necrosis in Fischer rats.Otolaryngol Head Neck Surg,1987,96:96-98.
2.Angel MF,Narayanan K,Swartz WM,et al.The etiologic role of free radicals in hematoma-induced flap necrosis.Plast Reconstr Surg,1986,77:795-803.
3.黎鳌,杨果儿,郭恩覃,主编.整形和烧伤手术学全集.北京:人民军医出版社,1996,439-442.
4.张振信,康淑擅,杨旅军,等.皮肤软组织扩张术并发症的教训.实用美容整形外科杂志,1998,4:61-63.
5.Bartell TH,Mustoe TA,Animal models of man tissue expansion.Plast.Reconstr.Surg.1989,83:681-686.
6.施新猷 主编.医学动物实验方法.北京:人民卫生出版社,1983,52-55.
7.Milton SH.The effects of "delay" on the survival of experimental pedicled skin flaps.Br J Plast Surg.1969 Jul;22:244-252.
8.Daniel RK,Kerrigan CL.The omnipotential pig buttock flap.Plast Reconstr Surg,1982,70:11-16.
9.陈铭,徐长军,秦建兵.等.红细胞对脑出血后脑水肿的影响及去铁胺的作用.中华神经外科疾病研究杂志,2005,4:265-266.
10.王京娥,张勇,丁新生,等.二异丙酚对脑出血大鼠脑组织NF-κB表达及超微结构的影响.中国脑血管病杂志,2007,4:172-176.
11.Karl-Heinz Diehl,Robin Hull,David Morton,et al.A Good Practice Guide to the Administration of Substances and Removal of Blood,Including Routes and Volumes.Journal of Applied Toxicology,2001,21,15-23.
12.王炜 主编.整形外科学.浙江科学技术出版社,1999,293-323.
13.Angel MF,Ramasastry SS,Swartz WM,et al.The critical relationship between free radicals and degrees of ischemia:evidence for tissue intolerance of marginal perfusion.Plast Reconstr Surg,1988,81:233-239.
14.Ward CG,Bullen JJ,Rogers HJ.Iron and infection:new developments and their implications.J Trauma.1996,41:356-64.
15.Hershko C.Iron chelators in medicine.Mol Aspects Med,1992,13:113-165.
16.Michael F,Feng Zhang,Barbara Rogers,et al.Role of Free Radicals in Necrosis of Skin Graft Compromised With Hematoma.2002,48:665-669.
17.Marlet JM,Barreto Fonseca JP,et al.Experimental determination of time of intracranial hemorrhage by spectrophotometric analysis of cerebrospinal fluid.J Forensic Sci,1982,27:880-888.
18.Matz.P,Weinstein PR,Sharp FR.Heme-oxygenase-1 and heat shock protein 70induction in glia and neurons throughout rat brain aider experimental intracerebral hemorrhage.Neurosurgery,1997,40:152-162.
19.钱忠明.铁代谢.基础与临床.北京:科学出版社,2000,293-305.
20.Hemnani T,Parihar MS.Reactive oxygen species and oxidative DNA damage.Indian J Physiol Pharmacol,1998,42:440-452.
21.Ferrari R,Agnoletti L,Comini L,et al.Oxidative stress during myocardial ischaemia and heart ailure.Eur Heart J,1998,19:2-11.
22.Bradley P,Priebat DA,Christensen RD,et al.Messurement of cutancous inflammation:estimation of neutrophil content with an enyme maker.J Invest Dermatol,1998,78:206-209.
1.Gelehrter TD,Sznycer-Laszyk R.Thrombin induction of plasminogen activator-inhibitor in cultured human endothelial cells.J Clin Invest,1986,77:165-169.
2.McNamara CA,Sarembock IJ,Gemple LW,et al.Thrombin stimulates proliferation of cultured rat aortic smooth muscle cells by a preoteolytically activated receptor.J Clin Invest,1993,91:94-98.
3.Fager G.Thrombin and proliferation of vascular smooth muscle cells.Circ Res,1995,77:645-650.
4.鲁开化,艾玉峰,郭树忠,主编.新编皮肽软组织扩张术.上海:第二军医大学出版社,2007,57-58.
5.Worseg A,Kuzbari R,Tairych G,et al.Long term results of inflatable mammary implants.Br J Plast Surg,1995,48:183-188.
6.Chen LB,Buchanan JM.Plasminogen-independent fibrinolysis by proteases produced by transformed chick embryo fibroblasts.Proc.Natl Acad Sci USA,1975,72:132-1136.
7.Carney DH,Cunningham DD.Cell surface action of thrombin is sufficient to initiate division of chick cells.Cell.1978,14:811-823.
8.Berk DC,Taubmanm MB,Griendling KK,et al.Thrombin-stimulated events in cultured vascular smooth-muscle cells.Biochem.J.1991,274:799-805.
9.魏东,陈文慧.羟脯氨酸含量的测定方法优化.现代中西医结合杂志,2005,14:2479-2480.
10.蔡文琴,主编.现代实用细胞与分子生物学实验技术.北京:人民军医出版社,2003:9-10.
11.杨志明,主编.组织工程学.北京:化学工业出版社,2002,155-158.
12.Coughlin SR.Thrombin signalling and protease-activated receptors.Nature,2000,407:258-264.
13.Tulinsky A.Molecular interactions of thrombin.Semin Thromb Hemost,1996,22:117-124.
14.Camerer E,Huang W,Coughlin SR.Tissue factor-α and factor X-dependent activation of protease-activated receptor 2 by factor VIIa.Proc Natl Acad Sci USA,2000,97:5255-5260.
15.Kaufmann R,Patt S,Zieger M,et al.The two-receptor system PAR-1/PAR-4mediates α-thrombin-induced[Ca~((2+))]immobilization in human astrocytoma cells.J Cancer Res Clin Oncol,2000,126:91-94.
16.Bartha K,Domotor E,Lanza F,et al.Identification of thrombin receptors in rat brain capillary endothelial cells.J Cereb Blood Flow Metab,2000,20:175-182.
17.Turgeon VL,Lloyd ED,Wang S,et al.Thrombin perturbs neurite outgrowth and induces apoptotic cell death in enriched chick spinal motoneuron cultures through caspase activation.J Neurosci,1998,18:6882-6291.
18.Rachel C,CHAMBERS,Karim DABBAGH,et al.Thrombin stimulates fibroblast procollagen production via proteolytic activation of protease-activated receptor 1.Biochem J,1998,333:121-127.
19.T.D.Hewitson,M.Martic,K.J.Kelynack,et al.Thrombin Is a Pro-Fibrotic Factor for Rat Renal Fibroblasts in vitro.Nephron Exp Nephrol,2005,101:e42-e49.
20.Berry CN,Crirardot C,Lecoffre C,et al.Effects of the synthetic thrombin inhibitor argatroban on fihrin-or clot-incorporated thrombin:comparison with heparin and recombinant hirudin.Thromb Haemost,1994,72:381-386.
21.司徒镇强,吴军正.细胞培养.第一版,西安:世界图书出版公司,2001,6-85.
22.Edwards CA,O' Brien WD.Modified assay for determination of hydroxyproline in a tissue hydrolyzate.Clin Chim Acta,1980,104:161-167.
23.Hewiston TD,Becker GJ.Interstitial myofibroblasts in IgA glomerulonephritis.Am.J.Nephrol.1995,15:111-117.
24.Zhang KY,and Phan S H.Inhibition of myofibroblast apoptosis by transforming growth factor beta.Am.J.Respir.Cell Mol.Biol,1999,21:658-665.
25.Zhang H,Gharaee-Kermani M,Zhang K,et al.Lung fibroblast alpha-smooth muscle actin expression and contractile phenotype in bleomycin-induced pulmonary fibrosis.Am.J.Pathol.1996,148:527-537.
26.Roy SG,Nozaki Y,Phan SH.Regulation of alpha-smooth muscle actin gene expression in myofibroblast differentiation from rat lung fibroblasts.Int J Biochem Cell Biol.2001,33:723-34.
27.Desmoulie re A,Rubbia-Brandt L,Abdiu A.,et al.α-Smooth muscle actin is expressed in a subpopulation of cultured and cloned fibroblasts.Exp Cell Res,1992,201:64-73.
28.Galina S Bogatkevich,Elena Tourkina,Richard M.Silver,et al.Thrombin Differentiates Normal Lung Fibroblasts to a Myofibroblast Phenotype via the Proteolytically Activated Receptor-1 and a Protein Kinase C-dependent Pathway.The Jouralof Biological Chemeistry,2001,276:45184-45192.
29.Mutch NJ,Robbie LA,Booth NA.Human thrombi an abundance of active thrombi n.Thromb H aemost,2001,86:1028-1034.
30.Bar-Shavit,R.,Kahn A,FentonII JW,et al.Chemotactic response of monocytes to thrombin.J Cell Biol,1983,96:282-285.
31.Bar-Shavit R,Khan A,FentonII,G D et al.Receptor-mediated chemotactic response of a macrophage cell line(J774) to thrombin.Lab.Invest.1983,49:702-707.
32.Bizios R,Lai L,Fenton II JW,et al.Thrombin-induced chemotaxis and aggregation of neutrophils.J Cell Physiol.1986,128:485-490.
33.吕宏开,朱庆生,王军.全自动显微镜及图象分析系统处理免疫组化图象.中国体视学与图象分析,2004,9:37-40
34.Bar-Shavit R,Sabbagh V,Lampugnani MG,et al.An Arg-Gly-Asp sequence within thrombin promotes endothelial cell adhesion.J Cell Biol,1991,112:335-344.
35.McNamara CA,Sarembock IJ,Gimple LW,et al.Thrombin stimulates proliferation of cultured rat aortic smooth muscle cells by a proteolytically activated receptor.J Clin Invest,1993,91:94-98.
36.White HD.Direct-thrombin inhibition and thrombolytic therapy:rationale for the Hirulog and Early Reperfusion Occlusion(HERO-2) Trial.AmJ Cardiol,1998,82: 57-62.
37.Bauer M,BrandsteLter H,Turk D,et al.Crystallographic determination of thrombin in complexes with small synthetic inhibitors as a starting point for the receptor-based design of antithrombin.Sem in Thromb Hemos,1993,19:352-360.
38.Hantgan RR,Jerome WG,Hursting MJ,et al.No effect of clot age or thrombolysis on argatroban's inhibition of thrombin,Blood,1998,92:2064-2074.
39.Hamada R,Matsuoka H._Antithrombin therapy for intracerebral hemorrhage.Stroke.2000,31:794-795.
40.Kitaoka T,Hua Y,Xi G,Hoff JT,Keep RF.Delayed argatroban treatment reduces edema in a rat model of intracerebral hemorrhage.Stroke.2002,33:3012-3018.
41.Matsuoka H,HamadaR.Role of thrombin in CNS damage associated with intracerebral haemorrhage:opportunity for pharmacological ntervention? CNS Drugs.2002,16:509-516.
42.Hantgan RR,Jerome WG,Hursting MJ.No effect of clot age or thrombolysis on argatroban's inhibition of thrombin.Blood,1998,92:2064-2070.
43.Pantoni L,Sarti C,Inzitari D,et al.Cytokines and cell adhesion molecules in cerebral ischemia:experimental bases and therapeutic perspectives.Arterioscler Thromb Vasc Biol,1998,18:503-513.
44.Xue M,DeI Bigio MR.Intracortical hemorrhage injury in rats:relationship between blood fraction and brain cell death.Stroke,2000,32:1721-1727.
45.Takagi K,Tsuchiya Y,Ohwaki K,et al.Body temperature in the super-acute phase of the cerebrovascular disorders:mainly focused on the subarachnoid hemorrhage.No To Shinkei.2002,54:139-45.
46.高长越,周华东,陈曼娥.凝血酶介导大鼠脑组织细胞间黏附分子-1表达与白细胞浸润的研究.中国危重病急救医学,2005,17:552-554.
47.关景霞,孙圣刚,曹学兵,等.脑血肿周围细胞间黏附分子-1mRNA表达及白 细胞浸润的实验研究.临床神经病学杂志,2004,17:432-434.
48.Bradley P,Priebat DA,Christensen RD,et al.Messurement of cutancous inflammation:estimation of neutrophil content with an enyme maker.J Invest Dermatol,1998,78:206-209.
49.Rothlein R,Dustin ML,Marlin SD,et al.A human intercellular adhesion molecule(ICAM-1) distinct from LFA-1.J Immunol,1986,137:1270-1274.
50.Wawryk SO,Novotny JR,Wicks IP,et al.The role of the LFA-1/ICAM-1interaction in human leukocyte homing and adhesion.Immunol Rev,1989,108:135-161.
51.Chirathaworn C,Kohlmeier JE,Tibbetts SA,et al.Stimulation through intercellular adhesion molecule-1 provides a second signal for T cell activation.J Immunol,2002,168:5530-5537.
52.Becker MD,Garman K,Whitcup SM,et al.Inhibition of leukocyte sticking and infiltration,but not rolling,by antibodies to ICAM-1 and LFA-1 in murine endotoxin-induced uveitis.Invest Ophthalmol Vis Sci,2001,42:2563-2566.
53.李江,鲁开化,艾玉峰,等.扩张囊外纤维包膜的组织学结构及其意义.中华医学美学美容杂,2001,7:191-192.
54.李江,鲁开化,艾玉峰,等.皮肤持续扩张后软组织的形态学改变.第四军医大学学报,1999,20:975.
55.李江,鲁开化,艾玉峰,等.扩张后皮肤软组织收缩机理的实验研究.中华整形外科杂志,2002,18:173-175.
56.Coleman D J,Sharpe DT,Naylor IL,et al.The role of the contractile fibroblast in the capsules around tissue expanders and implants.Br J Plast Surg.1993,46:547-56.
57.Madden JW,Morton D Jr,Peacock EE Jr.Contraction of experimental wounds.I.Inhibiting wound contraction by using a topical smooth muscle antagonist.Surgery.1974,76:8-15.
58.Rudolph R,Guber S,Suzuki M,et al.The life cycle of the myofibroblast.Surg Gynecol Obstet.1977,145:389-94.
59.Lee P,Squier CA,Bardach J.Enhancement of tissue expansion by anticontractile agents.Plast Reconstr Surg.1985,76:604-610.
60.Joseph J.The effects of theophylline and papaverine in enhancing skin expansion in guinea pigs.EurJ Plast Surg,1988,11:60-65.
61.张兆波,贾保祥.脑出血后凝血酶的毒性作用及抗凝血酶治疗现状.中华神经医学杂志,2006,5(10):1076-1078.
62.Lewis BE,Wallis DE,Berlkowitz SD,et al.Argatroban anticoagulant therapy in patients with heparin-induced thrombocytopenia.Circulation,2001,103:1838-1843.
63.Lcwis BE,Matthai WH Jr,Cohen M,et al.Argatroban anticoagulation during percutanceous coronary intervention in patients with with heparin-induced thrombocytopenia.Cathete Cardiovasc Interv,2002,57:177-184.
64.Ohteki H,Furukawa K,Ohnishi H,et al.Clinical experience of Argatroban for anticoagulation in cardiovascular surgery.Jpn J Thorac Cardiovasc Surg.2000,48:39-46.
1. Blair VP. The delayed transfer of long pedicle flaps in plastic surgery. Surg Gynecol Obstet 1921, 33, 261-92.
2. Polk HC, Miles AA. Enhancement of bacterial infection by ferris ion: kinetics, mechanisms, and surgical significance. Surgery 1971, 70:71-77.
3. Feng LU, Berger BE, Lysz TH, et al. Vasoactive prostaglandins in the impending no-reflow state: evidence for a primary disturbance in microvascular tone. Plast Reconstr Surg 1988, 81:755-767.
4. Mulliken JB, Healey NA. Pathogenesis of skin flap necrosis from underlying hematoma. Plast Reconstr Surg, 1979, 63: 540-545.
5. Hillelson RL, Glowacki J, Healey NA, et al. A microangiographic study of hematoma-associated flap necrosis and salvage with isoxsuprine. Plast Reconstr Surg, 1980, 66:528-533.
6. Angel MF, Haddad J, Abramson M. A free radical scavenger reduced hematoma-induced necrosis in Fischer rats. Otolaryngol Head Neck Surg, 1987, 96: 96-98.
7. Angel MF, Narayanan K, Swartz WM, et al. The etiologic role of free radicals in hematoma-induced flap necrosis. Plast Reconstr Surg, 1986, 77:795-803.
8. Smahel J. Development of hematoma under a free skin autograft. Plast Reconstr Surg, 1965,35:207-211.
9. Creech BJ, DeVito RV, Eade GG. Viability of split-skin grafts from pigs following incubation on autologous blood or serum for various periods. Plast Reconstr Surg, 1973, 51:572-574.
10. Michael F, Feng Zhang, Barbara Rogers, et al. Role of Free Radicals in Necrosis of Skin Graft Compromised With Hematoma. 2002, 48: 665-669.
11. Angel MF, Ramasastry SS, Swartz WM, et al. Free radicals: basic concepts concerning their chemistry, pathophysiology, and relevance to plastic surgery. Plast Reconstr Surg, 1987, 79: 990-997.
12. Aikens J, Dix TA. Prehydroxyl radical (HOO) initiated lipid peroxidation. The role of fatty acid hydroperoxides.J Biol Chem,1991,266:15091-15098.
13.Darr D,Fridovich I.Free radicals in cutaneous biology.J Invest Dermatol,1994,102:671-675.
14.Pang CY.Ischemia-induced reperfusion injury in muscle flaps:pathogenesis and major source of free radicals.J Reconstr Microsurg,1990,6:77-83.
15.Granger DN.Role of xanthine oxidase and granulocytes in ischemia-reperfusion injury.Am J Physiol,1988,255:H1269-H1275.
16.Angel MF,Ramasastry SS,Swartz WM,et al.The critical relationship between free radicals and degrees of ischemia:evidence for tissue intolerance of marginal perfusion.Plast Reconstr Surg,1988,81:233-239.
17.黎鳌,杨果凡,郭恩覃,主编.整形和烧伤手术学全集.北京:人民军医出版社,1996,439-442.
18.张振信,康淑擅,杨旅军,等.皮肤软组织扩张术并发症的教训.实用美容整形外科杂志,1998,4:61-63.
19.栾杰,张旭辉,陈莉,等.组织扩张过程中的感染及其影响因素分析.中华整形烧伤外科杂志,1998,3:137-139
20.鲁开化,艾玉峰,郭树忠,主编.新编皮肤软组织扩张术.上海:第二军医大学出版社.2007,57-58.
21.庄洪兴,罗家麟,廖杰,等.头面部皮肤软组织扩张术的并发症与防治.中华整形烧伤外科杂志,1995,11:455-456.
22.赵向东,沈江涌,高庆祥,等.皮肤软组织扩张术的并发症及其预防.宁夏医学院学报,2005,27:53-54.
23.鲁开化,郭树忠,艾玉峰,等.皮肤扩张术20年临床应用的回顾.中国实用整形美容外科杂志,2005,16:209-210.
24.王朝阳,朱世泽,刘祖民.负压引流在皮肤软组织扩张术中的应用.中华医学美学美容杂志,2001,7:186-187.
25.张超,王欢,刘春军,等.胶原蛋白海绵在皮肤扩张术中的应用.中国美容医学,2007,16:609-610.
26.Rees TD,Lee YC,Coburn RJ.Expanding hematoma after rhytidectomy:A retrospective study.Plast.Reconstr.Surg.,1973,51:149-153.
27. Grover R, Jones BM, Waterhouse N. The prevention of haematoma following rhytidectomy: A reviewof 1078 consecutive facelifts. Br J Plas Surg, 2001, 54: 481-486.
28. Rees TD, Barone CM., Valauri FA, et al. Hematomas requiring surgical evacuation following face lift surgery. Plast. Reconstr. Surg. 1994, 93: 1185-1190.
29. Baker DC, Stefani S, Chiu ES. Reducing the incidence of hematoma requiring surgical evacuationfoUowing male rhytidectomy: A 30-year review of 985 cases. Plast. Reconstr. Surg. Manuscript accepted for publication. Plast Reconstr Surg, 2005, 116: 1973-1987.
30. Barry M, Jones M, et al. Avoiding Hematoma in Cervicofacial Rhytidectomy: A Personal 8-Year Quest. Reviewing 910 Patients. Plast. Reconstr. Surg, 2004, 113: 381-387.
31. Lawson W, Naidu RK. The male facelift: An analysis of 115 cases. Arch. Otolaryngol. Head Neck Surg. 1993, 11: 535-451.
32. Grover R, Jones BM, Waterhouse N. The prevention of haematoma following rhytidectomy: A reviewof 1078 consecutive facelifts. Br. J. Plast. Surg. 2001, 54: 481-486.
33. Baker TJ, Gordon HL, Mosienko P. Rhytidectomy: a statistical analysis. Plast. Reconstr Surg. 1977, 59: 24-30.
34. Mayrovitz HN, Regan MB. Gender differences in facial skin blood perfusion during basal and heated conditions determined by laser Doppler flowmetry. Microvasc Res. 1993,45: 211-218.
35. Daniel C. Baker, M.D., William A, et al. Reducing the Incidence of Hematoma Requiring Surgical Evacuation following Male Rhytidectomy: A 30-Year Review of 985 Cases .Plast.Reconstr Surg. 2005, 116: 1973-1985.
36. Baker TM, Stuzin JM, Baker TJ, et al. What's new in plastic surgery. Clin Plast Surg, 1996,23:3-16.
37. Daniel C, Baker, M.D, Ernest S, et al. Bedside Treatment of Early Acute Rhytidectomy Hematomas. Plast. Reconstr. Surg, 2005, 115: 2119-2122.
38.Pitanguy I,Ceravolo MP.Hematoma post-rhytidectomy:How we treat it.Plast.Reconstr.Surg.1981.67:526-529.
39.王非可,侯秀英,张宝林.面部除皱术后血肿的预防和治疗.山西医药杂志.2001.30:354.
40.高岚,查元坤,李谆,等.双向引流在面部除皱术中的应用.中华医学美容杂志,1999,5:216-217.
41.Worseg A,Kuzbari R,Tairych G,et al.Long term results of inflatable mammary implants.Br J Plast Surg,1995,48:183-188.
42.Mladick RA."No-towch"submascular saline breast augmentation technique.Aesth Plast Surg,1993,17:183-192.
43.马显杰,鲁开化,艾玉峰,等.乳晕切口胸大肌后间隙隆乳纤维包膜挛缩的成因及防治.实用美容整形外科杂志,1999,10:225-226.
44.曾东,孙瑞霞,李勤,等.负压引流预防隆乳术后血肿形成和包膜挛缩的作用.广东医学,2002,23:42-43.
45.黄燮青,候明钟.负压引流在隆乳术中的应用.实用美容整形外科杂志.1998,10:241-242.
46.Jeffrey SS,Goodson WH Ⅲ,Ikeda DM,et al.Axillary lymphadenectomy for breast cancer without axillary drainage.Arch.Surg.1995,130:909-913.
47.Somers RG,Jablon LK,Kaplan MJ,et al.The use of closed suction drainage after lumpectomy and axillary node dissection for breast cancer:A prospective randomized trial.Ann.Surg.1992,215:146-149.
48.Colleen M.McCarthy,Joseph J.Disa,Andrea L.Pusic,M.D.The Effect of Closed-Suction Drains on the Incidence of Local Wound Complications following Tissue Expander/Implant Reconstruction:A Cohort Study.BREAST,2007,119:2018-2022.
49.Puttawibul P,Sangthong B,Maipang T,et al.Mastectomy without drain at pectoral area:A randomized controlled trial.J.Med.Assoc.Thai.2003,86:325-331.
50.Soon,PS,Clark J,Magarey CJ.Seroma formation after axillary lymphadenectomy with and without the use of drains.Breast,2005,14:103-107.
51.Mia Talmor,Thomas J,Fahey II,et al.Large volume liposuction complicated by retroperitoneal hemorrhage:management principles and implications for the quality improvement process.Plast Reconstr Surg,2000,105:2249-2250.
52.Pitman GH,Teimourian B.Suction Lipectomy complications and results by survey.Plas Reconst Surg.1985,76:65-72.
53.马越波,林君飞,戴峥.小吸管吸脂术减少手术并发症的探讨.中国美容医学,2000,9:49-50.
1.Xi Guohua,Keep RF,Hoff JT.Erythrocytes and delayed brain edema formation following intracerebral hemorrhage in rats.J Neurosurg,1998,89:991-996.
2.Marlet JM,Barreto Fonseca JP,et al.Experimental determination of time of intracranial hemorrhage by spectrophotometric analysis of cerebrospinal fluid.J Forensic Sci,1982,27:880-888.
3.Matz.P,Weinstein PR,Sharp FR.Heme-oxygenase-1 and heat shock protein 70induction in glia and neurons throughout rat brain after experimental intracerebral hemorrhage.Neurosurgery,1997,40:152-162.
4.LEE KEVIN R.Mechanisms of edema formation after intracerebral hemorrhage:effects of thrombin on cerebral blood flow,blood-brain barrier permeability,and cell survival in a rat model.J Neurosurg,1997,86:272-278.
5.Wu J,Hua Y,keep RF.Oxidative brain injury from extravasated erythrocytes after intracerebral hemorrhage.Brain Research,2002,953:45-52.
6.Simonni J,Feola M and Cani Zaro PC.Generation of free oxygen radicals and the toxicity of hemoglobin solutions,Biomater Artif cells.Artif Organs,1990,18:251-261.
7.Gutteridge J M.The antioxidant activity of haptoglobin towards heamoglobin stimulated lipid peroxidation.Biochim Biophys Acta,1987,917:219-223.
8.Regan RF,Panter SS.Hemoglobin potentiates excitotoxic injury in cortical cell culture.J Neurotrauma,1996,13:223-231.
9.Yip s,Ip Jk.Electrophysiological actions of hemoglobin on rat hippocampal CA1pyramidal neurons.Brain Res,1996,713:134-142.
10.Wu TW.The cytoprotective effect of Trolox demonstr-ated with three types of human cells.Cell Biol,1990,68:1189-1194.
11.何纲,刘茂才.中药防治自发性脑出血的实验研究进展.中药新药与临床药理,2002.13:54-58.
12.Macdonald RL.Weir BK.A review of hemoglobin and the pathogenesis of cerebral vasospasm.Stroke,1991,22:971-982.
13.Vollrath BA,Weir BK,Macdonald RL,et al.Intracellular mechanisms involved in the responses of cerebrovascular smooth-muscle cells to hemoglobin.J Neurosurg.1994,80:261-268.
14.Edwards DH,et al.The effect of chronic subarachnoid hemorrhage on basal endothelium derived relaxing factor activity in intrathecal cerebral arteries.Neurosurgery,1992,76:830-837.
15.Feng-Ping Huang,MD.Guo Hua Xi et al.Brain edema after experimental intracerebral hemorrhage:role of Hemoglobin degradation products.J Neurosurg,2002,2:287-293.
16.Guohua Xi,Ya Hua,R Rich Bhasin,BS.Effects of Extravasated Red blood cells on Blood Flow Blood-Brain Barrier Integrity.Stoke,2001,12:2933-2938.
17.Yang GY,Betz AL,Cherevert TL,et al.Experimental intra cerebral hemorrhage:relationship between brain edema,blood flow,and blood-brain-barrier permeability in rats.J Neurosurg,1994,81:93-102.
18.Regan RF,Panter SS.Neurotoxicity of hemoglobin in cortical cell culture.Neurosci Lett,1993,153:219-222.
19.贺双腾.脑溢安对血红蛋白损伤神经元神经生长因子和白细胞介素1-β表达的影响.湖南医科大学学报,2001,26:26-28.
20.Gonzales S,Erario MA,Tomar.Heme Oxygenase-1 induction and Dependent Increase in Ferritin a Protective antioxidant stratagem in hemin-treated rat brain.2002,24:161-168.
21.Hall NC,Packard BA,Hall NC.Protein oxidation and enzyme susceptibility in white and gray matter with in vitro oxidative stress:relevance to brain injury from intracerebral hemorrhage.Cell Mol Biol(Noisy-le-grand),2000,46:673-83.
22.Anderson DK,Means ED.Lipid peroxidation in spinal core Feclz induction and protection with antioxidants.Neurochem Pathol,1983,1:249-264.
23.Leclerc L,Marden M,Poyart C.Inhibition of erythrocyte Ca~(2+)/mg~(2+),ATP ase by nonheme iron.Biochim Biophys Acta.1991,1602:35-38.
24.Siesjo BK,Agardh CD,Bengtsson F.Free radicals and brain damage. Cerebrovasv Brain Metab,1989,1:165-211.
25.夏鹰,陈衍成等.脑出血.血肿周围脑组织的糖代谢,氧自由基及血一脑屏障的变化与脑水肿形成的关系,复旦学报,2002.29:119-121.
26.Peeling J,Yan HJ,chen SG,et al.Protective effects of free radical inhibitors in intracerebral hemorrhage in rat.Brain Res,1998,795:63-72.
27.Guohua Xi MD,Ya Hua,MD,R Rich Bhasin,BS.Erects of Extravasated Red blood cells on Blood Flow and Blood-Brain Barrier Integrity.Stoke,2001,12:2933-2938.
28.Verma A,Hirsh DJ,Glatt.Carbor monoxide:a putative neural messenger.Science,1994,259:381-384.
29.Tansen TW,Bratlid D,walaas SI.Bilirubin decrease phosphorylation of synapsin I,a synaptic vesicle-asociated neuronal phosphoprotein in intract synaptosomes from rat cerebral cortex.Pediatr Res,1988,23:219-223.
30.Lves Nk,Bolas NM.The effect of bilirubin on brain energy metabolism during hyperosmolar opening of the BBB:an in vivo study using 3 lp nuclear magnetic resonance spectroscopy.Pediatr Res,1989,26:356-361.
31.Amit Y,Chan G.Bilirubin toxicity in a neuroblastoma cell line and mitochondrial function.Pediatr Res,1989,25:364-368.
32.Amit Y,Brenner T.Age-dependent sensitivity of cultured rat glial cells to bilirubin toxicity.Exp Neurol,1993,121:248-255.
33.Di Cera E,Cantwell AM.Determinants of thrombin specificity.Ann N Y Acad Sci.2001,936:133-146.
34.Ayala YM,Cantwell AM,Rose T,Bush LA,Arosio D,Di Cera E.Molecular mapping of thrombin-receptor interactions.Proteins.2001,45:107-116.
35.Kaufmann R,Patt S,Zieger M,et al.The two-receptor system PAR-1/PAR-4mediates alpha-thrombin-induced[Ca(2+)](i) mobilization in human astrocytoma cells.J Cancer Res Clin Oncol.2000,126:91-94.
36.Xi G,Keep R,Hoff J.Mechanisms of brain injury after intracerebral hemorrhage.Lancet Neurol.2006,5:53-63.
37.Xi G,Wagner KR,Keep RF.et al.The role of blood clot formation on early edema development following experimental intracerebral hemorrhage. Stroke. 1998,29:2580-2586.
38. Bartha K, Domotor E, Lanza F, et al. Identification of thrombin receptors in rat brain capillary endothelial cells. J Cereb Blood Flow Metab. 2000, 20:175-182.
39. NagyZ, Kolev K, Csonka E, et al. Contraction of human brain endothelial cells induced by thrombogenic and fibrinolytic factors. An in vitro cell culture model. Stroke, 1995, 26: 265-270.
40. Lee KR, Kawai N, Kim S, et al. Mechanisms of edema formation after intracerebral hemorrhage: effects of thrombin on cerebral blood flow, blood-brain barrier permeability, and cell survival in a rat model. J Neurosurg, 1997, 86: 272-278.
41. Jiang Y, Wu J, Hua Y. et al. Thrombin receptor activation and thrombin-induced brain tolerance. J Cereb Blood Flow Metab, 2002, 22: 404-410.
42. Gong C, BouIts N, Qian J, et al. Intracerebral hemorrhage-induced neuronal death. Neurosurgery, 2001, 48: 875-882.
43. Donovan FM, Pike CJ, Cotman CW, et al. Thrombin induces apoptosis cultured neurons and astrocytes via a pathway requiring tyrosine kinase and RhoA activities. J Neurosci, 1997, 17: 5316-5326.
44. Jerius H, Beall A, Woodrum D, et al. Thombin-induced vasospasm: cellular signaling mechanisms. Surgery, 1998, 123: 46-50.
45. Hua y, Schallert T, Keep RF, et al. Behavioral tests after intracerebral hemorrhage in the rat. Stroke. 2002, 33: 2478-2484.
46. Matsuoka H, Hamada R. Role of thrombin in CNS damage associated with intracerebral haemorhage: opportunity for pharmacological intervention? CNS Drugs, 2002, 16: 509-516.
47. Xi G, Hua Y, Keep RF, et al. Induction of coIligin may attenuate brain edema following intracerebral hemorrhage. Acta Neurochir Suppl, 2000, 76: 501-505.
48. Xi G, Hua Y, Keep RF, et al. Activation of p44/42 mitogen activated protein kinases in thrombin-induced brain tolerance. Brain Res, 2001, 895:153-159.
49. Masada T, Hua Y, Xi G, et al. Attenuation of intracerebral hemorrhage and thrombin-induced brain edema by overexpression of interleukin-1 receptor antagonist. J Neurosurg, 2001, 95: 680-686.
50. Hua Y, Wu J, Keep R. et al. Tumor necrosis factor-alpha increases in the brain following intracerebral hemorrhage and thrombin stimulation. Neurosurgery. 2006, 58:542-550.
51. Nguyen M, Arkell J, Jackson CJ. Thrombin rapidly and efficiently activates gelatinase a in human microvascular endothelial cells via a mechanism independent of active mtl matrix metalloproteinase. Lab Invest. 1999, 79: 467-475.
52. Xi G, Keep RF. Attenuation o fthrombin-induced brain edema by cerebral thrombin preconditioning. Stroke, 1999, 30: 1247-1255.
53. Masada T, X I G, Hua Y, et al. The effects of thrombin preconditioning on focal cerebral ischemia in rats. Brain Res, 2000, 867: 173-179.
54. Jiang Y, Wu J, Hua Y, et al. Thrombin-receptor activation and thrombin-induced brain tolerance. J Cereb Blood Flow Metab, 2002, 22: 404-410.
55. Landau E, Tirosh R, Pinson A, et al. Protection of thrombin receptor expression under hypoxia. J Biol Chem, 2000, 275: 2281-2287.
56. Hamada R, Matsuoka H. Antithrombin therapy for intracerebral hemorrhage. Stroke. 2000, 31:794-795.
57. Kitaoka T, Hua Y, Xi G, et al. Delayed argatroban treatment reduces edema in a rat model of intracerebral hemorrhage. Stroke. 2002, 33: 3012-3018.
58. Matsuoka H, HamadaR. Role of thrombin in CNS damage associated with intracerebral haemorrhage: opportunity for pharmacological ntervention? CNS Drugs. 2002, 16: 509-516.