云南白药及主要成分骨形成作用的实验研究
|
摘要
研究背景：
云南白药的主要组成成分有三七、重楼、麝香等名贵中草药,对于骨伤治疗有其独特的疗效,但对其促进骨折愈合的作用机理至今尚无明确的生物学机理解释。
目前,有关白药治疗骨折的许多研究仍停留在临床疗效观察阶段,对于白药复方的有效成分的分析及作用机理却很少见报道。由于云南白药的组成配方是保密的原因,有关白药复方的药理、药效研究难以深入,更缺乏从分子、细胞水平上进行深入系统的研究,缺少良好的现代病理生理学基础。同时,由于药理研究模式基本上沿用西药药理学的模型,不能客观地反映白药复方的主要药效,无法切实表达复方的临床疗效,这也是白药复方研究能否突破低水平重复的关键。
研究目的：
作为我国著名中成药-云南白药,有必要通过体外细胞培养方法以及进一步的动物实验研究,采用现代分子生物学技术对其主要成分对骨折愈合过程中的成骨作用和作用机理进行深入研究,进一步揭示云南白药对骨组织愈合修复作用的分子机制。
研究方法：
本课题设计首先通过拆方研究,观察云南白药及十种组成成分(代号)对体外鼠骨髓基质细胞分化、增殖以及对骨基质蛋白及相关生长因子表达变化的影响,阐明云南白药各原药成分在MSC成骨诱导分化中的作用、促进骨折修复的细胞、分子的作用机理。
在此基础上,从前期实验中寻找出高效低毒成分,将分析和综合研究有机结合起来,进行重新配伍组方,并将新配伍局部应用到兔骨折愈合过程中,通过动态观察骨折修复的情况,x线检测,组织学、以及免疫组化等分子生物学等方法,观察重组配伍的云南白药对骨组织修复过程中几种细胞因子及胶原的影响,明确云南白药复方制剂中各原药成分所处的地位和作用,进一步探讨云南白药促进骨折愈合的作用机制。
研究结果：
1.云南白药的多种成分通过促进鼠骨髓基质细胞产生BMP和TGF-β等因子,从而诱导MSCs向成骨细胞转化,促进成骨细胞活性,同时促进骨基质合成,其中云南白药E、G、H、I、J五种原药成分细胞毒性较小,促进基质细胞转化作用较明显。
2.在实验性兔骨折愈合的早期、中期阶段,1到6号组方白药能促进内皮细胞增殖,诱导新生血管的形成,从而促进骨折的愈合过程。
3.本研究中还发现云南白药还通过促进Ⅰ型胶原与其受体整合素αβ结合,激活FAK途径,促进成骨细胞的增殖和分化,并具有促进局部血管生成,改善局部供氧条件的作用,这可能有助于抑制骨折愈合过程中的软骨化骨,促进膜内成骨以加速新骨形成的速度和质量,缩短骨折愈合的过程。
研究结论：
1.骨折愈合过程早、中期,云南白药主要通过直接影响及间接影响促进骨髓基质细胞向成骨细胞分化,促进成骨细胞活性,诱导骨折周围组织新血管生成,引起骨折局部血供丰富及细胞活跃等机制,从而加快骨折的愈合。
2.云南白药的多种成分中可能具有某些类似骨生长因子和血管生长因子的作用,其中E、G、H、I、J五种原药成分细胞毒性较小,促进骨髓基质细胞转化作用较明显,具有进一步的研究开发基础。
3.白药单个原药成分之间、E、G、H、I、J五种原药成分重新组方及白药复方的药效学比较实验结果较为复杂。新组方白药对骨折愈合的促进作用虽然是共同的,但各新组方之间却没有显示出明显的相关性及差异性,白药中各单个原药成分对复方药效起不起作用、起多大作用,其具体有效成分,尤其是白药复方制剂的有效成分和作用机制尚待进一步深人研究。
Background
The essential component of the famous "Yunnan Baiyao" are san qi (Panax notoginseng) and other Chinese Herbal Medicine, and it is already used widely in clinical almost one hundred years, and has unique, reliable curative effect especially in treating of bone fracture. But we know less about the biological Mechanism of action.
At present, a lot of studies about Baiyao are stay at the observing stage of clinical effects, rarely study about the analysis of effective constituent and mechanism of action. And the study about pharmacology and pesticide effect of Compound Baiyao is Low levels and repeated. And short of systematic and Intensive study at molecular and cellular level, and lake of Good and modern basis of Pathophysiology.
The Pathophysiologic study model is practically the same as Western medicine. So is unable to reflect principal clinical and pesticide effects of compound Baiyao objectively, and this is strategic point of to can make a breakthrough an indicator of at the study of compound Baiyao.
Objective
As our country Famous "Yunnan baiyao", it is necessary to intensive study the bone formation and mechanism of Yunnan Baiyao and major components on treating bone fracture by means of making external cells (MSCs) cultivation and further animal experiment with adopting the contemporary Molecular biology technique, to explore the mechanism of actions of Yunnan Baiyao and major components on promoting bone tissue repair.
Methods
This project, first, put a broken prescriptions study into practice by observing the Yunnan Baiyao and ten major components (code name) expect that proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) from
SD rats and the express change of bone Matrix Protein and related growth factor. To explore the effect of Yunnan Baiyao and each component on that proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and the cellular molecule mechanism of promoting the healing of bone fracture.
Second, to look for efficient and harmfulless component from earlier studies, and design the new compound Baiyao application locally in the healing of rabbit bone fracture by fall together analyse and comprehensive. To clear and definite the located position and action of Yunnan Baiyao and each component, and explore the mechanism of promoting the healing of bone fracture through dynamically observe the situation of bone tissue repairing by detection of x-Ray, histology, immunohistochemistry, molecular biology and the effect of Yunnan Baiyao and each component on some kinds of cell factor and collagen during the repairing of bone fracture.
Results
1. Variety of Yunnan Baiyao components stimulate the differentiation of osteoblasts derived from bone marrow stromal dells by that facilitate the production of some factors such as TGF-β,BMP, and encourage the activity of osteoblast, advance the composition of bone matrix. E, G, H, I, J components have obvious effect on the differentiation of bone marrow stromal dells and have lower cytotoxicity in all of Yunnan components.
2.One to six number new formulating Baiyao has obvious effect on promoting the healing of bone fracture by proliferation of endothelial cell and formation of new blood vessels in early and medium term during the repairing of rabbit bone fracture.
3.This study find that Baiyao also have obvious effect on proliferation and differentiation of osteoblast by promoting combination of collagen I and integrainαβand activating FAK-way and formatiing local new blood vessels and improving condition of local oxygen supply. It is possible contribute to accelarte speed and quality and reduce the process of the healing of bone fracture by that new formulating Baiyao suppresed the process of cartilagenous and promoted that of intramembranous ossification.
Conclusions
1. In early and medium term during the repairing of rabbit bone fracture, the possible mechanism of Yunnan Baiyao and major components in promoting bone healing was to directly or indirectly effect on the proliferation of BMSCs, enhance the activity of osteoblast, and the formation of new blood vessels, therefore, lead to blood supply plentiful and cell vigorous of fracture local.
2. It is possible the Yunnan Baiyao and major components had same stimulative fouction similar as bone growth factor and vascular growth factor. Among all of components, E、G、H、I、J had obvious stimulative effect on the differentiation of bone marrow stromal dells and have lower cytotoxicity, which had foundation of farther research and development.
3. The experimental result that the Compare pharmacological action of in each Baiyao component, new formulating Baiyao and compound Baiyao is complicated. Although the 1 to 6 number new formulating Baiyao has a common effect on promoting the healing of bone fracture, but this research did not show obvious relativity and Differentiation both between and within new formulating Baiyao components as we thought before. We did not understand that each Baiyao component play what kind of role in the compound Baiyao, and what is the specific active constituent in all of Baiyao components. Especially the definite constituent in which and the possible mechanism of promoting the healing of bone fracture need more intensive work to do.
云南白药的主要组成成分有三七、重楼、麝香等名贵中草药,对于骨伤治疗有其独特的疗效,但对其促进骨折愈合的作用机理至今尚无明确的生物学机理解释。
目前,有关白药治疗骨折的许多研究仍停留在临床疗效观察阶段,对于白药复方的有效成分的分析及作用机理却很少见报道。由于云南白药的组成配方是保密的原因,有关白药复方的药理、药效研究难以深入,更缺乏从分子、细胞水平上进行深入系统的研究,缺少良好的现代病理生理学基础。同时,由于药理研究模式基本上沿用西药药理学的模型,不能客观地反映白药复方的主要药效,无法切实表达复方的临床疗效,这也是白药复方研究能否突破低水平重复的关键。
研究目的：
作为我国著名中成药-云南白药,有必要通过体外细胞培养方法以及进一步的动物实验研究,采用现代分子生物学技术对其主要成分对骨折愈合过程中的成骨作用和作用机理进行深入研究,进一步揭示云南白药对骨组织愈合修复作用的分子机制。
研究方法：
本课题设计首先通过拆方研究,观察云南白药及十种组成成分(代号)对体外鼠骨髓基质细胞分化、增殖以及对骨基质蛋白及相关生长因子表达变化的影响,阐明云南白药各原药成分在MSC成骨诱导分化中的作用、促进骨折修复的细胞、分子的作用机理。
在此基础上,从前期实验中寻找出高效低毒成分,将分析和综合研究有机结合起来,进行重新配伍组方,并将新配伍局部应用到兔骨折愈合过程中,通过动态观察骨折修复的情况,x线检测,组织学、以及免疫组化等分子生物学等方法,观察重组配伍的云南白药对骨组织修复过程中几种细胞因子及胶原的影响,明确云南白药复方制剂中各原药成分所处的地位和作用,进一步探讨云南白药促进骨折愈合的作用机制。
研究结果：
1.云南白药的多种成分通过促进鼠骨髓基质细胞产生BMP和TGF-β等因子,从而诱导MSCs向成骨细胞转化,促进成骨细胞活性,同时促进骨基质合成,其中云南白药E、G、H、I、J五种原药成分细胞毒性较小,促进基质细胞转化作用较明显。
2.在实验性兔骨折愈合的早期、中期阶段,1到6号组方白药能促进内皮细胞增殖,诱导新生血管的形成,从而促进骨折的愈合过程。
3.本研究中还发现云南白药还通过促进Ⅰ型胶原与其受体整合素αβ结合,激活FAK途径,促进成骨细胞的增殖和分化,并具有促进局部血管生成,改善局部供氧条件的作用,这可能有助于抑制骨折愈合过程中的软骨化骨,促进膜内成骨以加速新骨形成的速度和质量,缩短骨折愈合的过程。
研究结论：
1.骨折愈合过程早、中期,云南白药主要通过直接影响及间接影响促进骨髓基质细胞向成骨细胞分化,促进成骨细胞活性,诱导骨折周围组织新血管生成,引起骨折局部血供丰富及细胞活跃等机制,从而加快骨折的愈合。
2.云南白药的多种成分中可能具有某些类似骨生长因子和血管生长因子的作用,其中E、G、H、I、J五种原药成分细胞毒性较小,促进骨髓基质细胞转化作用较明显,具有进一步的研究开发基础。
3.白药单个原药成分之间、E、G、H、I、J五种原药成分重新组方及白药复方的药效学比较实验结果较为复杂。新组方白药对骨折愈合的促进作用虽然是共同的,但各新组方之间却没有显示出明显的相关性及差异性,白药中各单个原药成分对复方药效起不起作用、起多大作用,其具体有效成分,尤其是白药复方制剂的有效成分和作用机制尚待进一步深人研究。
Background
The essential component of the famous "Yunnan Baiyao" are san qi (Panax notoginseng) and other Chinese Herbal Medicine, and it is already used widely in clinical almost one hundred years, and has unique, reliable curative effect especially in treating of bone fracture. But we know less about the biological Mechanism of action.
At present, a lot of studies about Baiyao are stay at the observing stage of clinical effects, rarely study about the analysis of effective constituent and mechanism of action. And the study about pharmacology and pesticide effect of Compound Baiyao is Low levels and repeated. And short of systematic and Intensive study at molecular and cellular level, and lake of Good and modern basis of Pathophysiology.
The Pathophysiologic study model is practically the same as Western medicine. So is unable to reflect principal clinical and pesticide effects of compound Baiyao objectively, and this is strategic point of to can make a breakthrough an indicator of at the study of compound Baiyao.
Objective
As our country Famous "Yunnan baiyao", it is necessary to intensive study the bone formation and mechanism of Yunnan Baiyao and major components on treating bone fracture by means of making external cells (MSCs) cultivation and further animal experiment with adopting the contemporary Molecular biology technique, to explore the mechanism of actions of Yunnan Baiyao and major components on promoting bone tissue repair.
Methods
This project, first, put a broken prescriptions study into practice by observing the Yunnan Baiyao and ten major components (code name) expect that proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) from
SD rats and the express change of bone Matrix Protein and related growth factor. To explore the effect of Yunnan Baiyao and each component on that proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells and the cellular molecule mechanism of promoting the healing of bone fracture.
Second, to look for efficient and harmfulless component from earlier studies, and design the new compound Baiyao application locally in the healing of rabbit bone fracture by fall together analyse and comprehensive. To clear and definite the located position and action of Yunnan Baiyao and each component, and explore the mechanism of promoting the healing of bone fracture through dynamically observe the situation of bone tissue repairing by detection of x-Ray, histology, immunohistochemistry, molecular biology and the effect of Yunnan Baiyao and each component on some kinds of cell factor and collagen during the repairing of bone fracture.
Results
1. Variety of Yunnan Baiyao components stimulate the differentiation of osteoblasts derived from bone marrow stromal dells by that facilitate the production of some factors such as TGF-β,BMP, and encourage the activity of osteoblast, advance the composition of bone matrix. E, G, H, I, J components have obvious effect on the differentiation of bone marrow stromal dells and have lower cytotoxicity in all of Yunnan components.
2.One to six number new formulating Baiyao has obvious effect on promoting the healing of bone fracture by proliferation of endothelial cell and formation of new blood vessels in early and medium term during the repairing of rabbit bone fracture.
3.This study find that Baiyao also have obvious effect on proliferation and differentiation of osteoblast by promoting combination of collagen I and integrainαβand activating FAK-way and formatiing local new blood vessels and improving condition of local oxygen supply. It is possible contribute to accelarte speed and quality and reduce the process of the healing of bone fracture by that new formulating Baiyao suppresed the process of cartilagenous and promoted that of intramembranous ossification.
Conclusions
1. In early and medium term during the repairing of rabbit bone fracture, the possible mechanism of Yunnan Baiyao and major components in promoting bone healing was to directly or indirectly effect on the proliferation of BMSCs, enhance the activity of osteoblast, and the formation of new blood vessels, therefore, lead to blood supply plentiful and cell vigorous of fracture local.
2. It is possible the Yunnan Baiyao and major components had same stimulative fouction similar as bone growth factor and vascular growth factor. Among all of components, E、G、H、I、J had obvious stimulative effect on the differentiation of bone marrow stromal dells and have lower cytotoxicity, which had foundation of farther research and development.
3. The experimental result that the Compare pharmacological action of in each Baiyao component, new formulating Baiyao and compound Baiyao is complicated. Although the 1 to 6 number new formulating Baiyao has a common effect on promoting the healing of bone fracture, but this research did not show obvious relativity and Differentiation both between and within new formulating Baiyao components as we thought before. We did not understand that each Baiyao component play what kind of role in the compound Baiyao, and what is the specific active constituent in all of Baiyao components. Especially the definite constituent in which and the possible mechanism of promoting the healing of bone fracture need more intensive work to do.
引文
[1]申海明.近30年来云南白药应用状况[J].云南医药杂志,1986,7(4)：46.
[2]潘振球等.单味云南白药外敷的应用概况.中国临床医生.2001；29(7)：49.
[3]朱大忠等.云南白药的药理及临床研究进展.中西医结合杂志.1987：7(6)：377～379.
[4]蔡锡麟等.云南白药的药理研究.中成药研究.1982：(8)： 37.
[5]孔令义,杨智,闵知大.对中药现代化中有效成分研究的思考.中草药,1998,29(5)：354～355.
[6]刘建立.中药复方疗效的物质基础及其化学成分研究方法.中成药,1992,14(10)：49.
[7]王永炎,赵宜军.从大黄复方的运用浅谈中药复方研究的方法与思路.中国中药杂志,2000,25(3)：131～132.
[8]何祥久,邱峰,姚新生.中药复方研究现状和思路.化学进展,2001,13(6)：481～484.
[9]罗国安,王义明.中药复方物质基础和药效相关性研究,世界学技术-中药现代化,1999,1：11～15.
[10]武孔云,梁光义,靳风云,等.中药复方药效物质基础研究的思路及方法.世界科学技术-中医药现代化,2003,5(6)：13～15.
[11]孙明瑜,谢鸣.试论证候与复方的研究思路.中国医药学报,2002,17(4)：196～198.
[12]姚新生,胡柯.中药复方的现代化研究.化学进展。1999,11(2)：192～196.
[13]程永现,赵淑敏.中药复方研究中的几个关键性问题-中医现代化思路探索.中国医药学报,2001,16(1)：63-65.
[14]徐东铭,徐雅红.中药复方药效活性成分研究思路初探.中国医药学报,2001,16(6)：52-54.
[15]刘良.中药复方基础研究的思路与方法探讨.中国实验方剂学杂志,2000,6(4)：1～4.
[16]方文贤.中药新药研究开发与思路创新的再思考.中国医药学报,2003,18(6)：365～369.
[17]李澎涛,乔延江,王永炎,等.对中药复方研究的思考.北京中医药大学学报,1999,22(5)：15～18.
[1]王和鸣,王力,李楠.巴戟天对骨髓基质细胞向成骨细胞分化影响的实验研究.福建中医学院学报.2004,14(3)：16～20.
[2]王银喜,王金熙,徐智敏,等.局部植入接骨中药提取物促进骨愈合的实验研究.中国骨伤,2003；16(6)：346～349.
[3]王斌,罗毅文,胡年宏.骨康方对大鼠MSCs体外向成骨细胞分化和ALP的影响.中国中医骨伤科杂志.2007,15(12)：5～9.
[4]韩立民,刘波,徐彭.淫羊藿对成骨细胞增殖的血清药理学研究.中医药学刊,2003,21(5)：678～680.
[5]祝晓玲,祝彼得.黄芪体外作用对贫血小鼠骨髓基质细胞分泌SCF的影响.细胞与分子免疫学杂志,2002,18(4)：396.
[6]Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precurors in normal and irradiated mouse hematopoietic organs. Exp Hematol,1976,4(5):267～74.
[7]Caplan AL. The mesengenic process. Clin Plast Surg,1994,21:429～435.
[8]Pitlenger MF, Mackay AM, Beck SC et al. Multilineage potential of adult human mesenchymal stem cells. Science,1999,284:143～147.
[9]Deans RJ, Moseley AB. Mesenchymal stem cells:Biology and potential clinical uses. Exp Hematol,2000,28:875～884.
[10]Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potennial of adult human mesenchymal stem cells. Science,1999,284:143～147.
[11]Nathanson MA. Bone matrix-directed chondrogenesis of muscle in vitro. J Clin Orthop, 1985,200:142～158.
[12]Nakahara H, Dennis J E, Bruder SP, et al. In vitro differentiation of bone and hypertrophic cartilage from periosteal-derived cells. Exp Cell Res,1991,195:492～503.
[13]Campagnoli C, Roberts IA, Kumar S, et al. Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood,2001,98:2396～ 2402.
[14]Chen TL, Shen WJ, Kraemer FB. Human BMP-7/0P-1 induces the growth and differentiation Of adipocytes and osteoblasts in bone marrow stromal cell cultures. Cell Biochem.2001, 82(2):187.
[15]支会英,李恩.补肾方药对去卵巢骨质疏松的骨组织ER表达的影响.中国中医基础医 学杂志,2001,7(4)：41～44.
[16]Rhee ST, EI-Bassiony L, Buchman SR, et al. Extracellular signal-related kinase and bone morphogenetic protein expression during distraction osteogenesis of the manndible:in vivo evidence of a mechanotransduction mechanism for differentiation and osteogenesis by mesenchymal precursor cells. Plast Reconstr Surg,2006,117(7):2243.
[17]郝瑞锋.申长虹.骨髓问充质干细胞向成骨细胞诱导的研究现状.天津医科大学学报。2007,13(3)：129～131.
[18]RINGE J, KAPS C, BURMESTER G R, et al. Stem cells for re—generative medicine: advances in the engineering of tissues and organs(Review). Naturwissenschaften,2002, 89(8):38-351.
[19]CLARKE D, FRISEN J. Differentiation potential of adult stem cells. Curr Opin Genet.2001, 11(5):575-80.
[20]Ross R, Raines EW, et al. The biology of platelet derived growth factor. Cell,1986,46:115.
[21]Robey PG, Yong MF, et al. Osteo-blasts synthesize and respond to transforming growth factor beta in vitro. J Cell Biol,1987,105:457.
[22]Andrew JW, Hoyland J, et al. Demonstraction of BMP2 mRNA expression in human fracture callus. J Bone Joint Surg,1993,75:(supp3),88.
[23]Cook-SD, Salkeld-SL, Brinker-MR, et al. Use of an osteoinductive biomaterial (rhOP-1) in healing large segmental bone defects. J Orthop-Trauma.1998,8; 12(6):407～412.
[24]DAVID C, REINER C, CARLA M, et al. Rapid expansion of recycling stem cells in cultures of plastic adherent cells from human bone marrow. Pronc Natl Acad Sci,2000,97(7):3213～ 3218.
[25]Brian Jonstone, Tomas M. In vitro Chondrogenesis of bone marrow-derived mesenchymal Progenitor cells. Experiment Cell Resarch,1998,238:265～271.
[26]夏万尧,商庆新,崔磊,等.骨髓间充质干细胞向软骨细胞表型定向诱导分化的实验研究[J].中华整形外科杂志,2002,18：12～14.
[27]Noble BS, Dean V, loueridge N. Dextran sulfate promotes the rapid aggregation of porcine bone-marrow stromal cells. Bone,1995,1(4):375～382.
[28]杨志明.组织工程基础与临床成都[M].成都：四川科学技术出版社,2000,37～57.
[29]Long MW, Robinson JA, A shcraft EA, et al. Regulation of human bone marrow-derived osteoprogenitor cells by osteogenic growth factors. J Clin Invest,1995,95(2):881.
[30]王蓓,汪维伟.问充质干细胞的研究进展[J].国外医学：外科学分册,2005,32(6)： 459～462.
[31]Turksen K, Aubin J E. Positive and negative immunoselection for enrichment of two classes of osteoprogenitor cells. J Cell Biol,1991,114:373～384.
[32]Maniatopoulos C, Sodek J, MelcherAH. Bone formation in vitro by stromal cells obtained from bonemarrow of young adult rats. Cell Tissue Res,1998,254:317.
[33]McQillan DJ, Richardson MD, Bateman JF. Matrix deposition by a calcifying human osteogenic sarcoma cell line (SAOS22). Bone,1995,16:415～426.
[34]Otsuka E, Yamaguchi A, Shigehisa H, et al. Characterizations of osteoblastic differentiation of stromal cell line ST2 that is induced by ascorbicacid. Am J Physiol,1999,277:132～138.
[35]Masaki I, Takenbu K, Shuichi A, et al. Bone morphogenetic protein-12 and-13 inhibit terminal differentiation of myoblasts, but do not induce their differentiation into osteoblast. Biochem Biophys Res Commun,1996,222:317～322.
[36]Kim KJ, Itoh T, Kotake S. Effects of recombinant human bone morphogenetic protein-2 on human bone marrow cells culture with various biomaterial. Biomed Mater Res,1997,35: 279～285.
[37]Gronthos S, Zannetion AC, Hay SJ, et al. Molecular and celluer characterization of highly purified stromal stem cells derived from human bone marrow. J Cell Sci,2003,116:1827～ 1835.
[38]Tsuda H, Wada T, Ito Y, et al. Efficient BMP2 gene transfer and bone formation of mesenchymal stem cells by a fiber-mutant adenoviral vector. J Mol Ther,2003,7:354～365.
[39]Rickard Dj, Sullivan TA, Shenker BJ, et al. Induction of rapid osteoblast differentiation in rat bone marrow stromal cell culture by dexamethasone and BMP-2. J Dev Biol,1994,161: 218～228.
[40]杨丕斌,胡侦明,劳汉昌.雌激素对骨折愈合过程中骨形态发生蛋白-4 mRNA表达的影响.中国矫形外科杂志,2003,5：620～621.
[41]Zhenming Hu, Sean A.F. Peel, George K. B. Sander, Cameron M. L. Clokie. The osteoinductive activity of bone morphogenetic protein (BMP) purified by repeated extracts of bovine bone. Growth Factors,2004,2:1～5.
[42]Rosier RN O, Keefe RJ, Hicks DG. The potential role of transforming growth factor beta in fracture healing. Clin Orthop.1998, (355 Suppl):294～300.
[1]柴本甫.骨折模型[J].中华外科杂志.1962.1(5)：299-301.
[2]李人杰,柳用墨,王琮武.旋磁场促进骨折愈合的初步实验研究.中华骨科杂志,1998,8(6)：445～447.
[3]Kraan MC, Smith MD, Weedon H et al. Measurement of cytokine and adhesion molecule expression in synovial tissue by digital image analysis. Ann Rheum Dis,2001;60; 296～298.
[4]吕翔.病理图像定量分析及其测量误差的控制.中国体视学与图像分析.2002,7(1)：58～62.
[5]Frost HM. Magovem CJ, Mack CA, et al. The biology of fracture healing. an overview for clinicians. Clin Orthop 1992:248(11):294.
[6]Arobson J. Cassnano C. Rowhim M, et al. Experimental and clinical expcrience with distraction osteogenesis. ClEfipalate Craniofac 1995:31(6):473.
[7]Eppley BL, Connolly DT, Winkelmann T, et al. Free bone graft reconstruction of irradiated facial tissue:Experimental effects of basic fibroblast growth factor stimulation. Plast Reconstr Surg 1996:88(1):1.
[8]Trueta P, Sacehi N, Lei J, et al. Single-step method of RNA is olation by acid guanidinium thiocyanate—phen01. chloroform extraction. Anal Biochem 1997; 162(1):156.
[9]Chuanyong Lu, Erik Hansen, Anna Sapozhnikova,et al.Effect of Age on Vascularization during Fracture Repair.J Orthop Res.2008,26:1384-1389.
[10]Ru Li, Duncan J. Stewart, Herbert P,et al.Effect of Cell-Based VEGF Gene Therapy on Healing of a Segmental Bone Defect.J Orthop Res.2009,27:8-14.
[11]FelTara N. Davis Smyth T. Vincenti V, et al. The biology of vascular endothelial growth factor. Endocr Res 1997:18(8):4.
[12]刘杰,孙正义,曹蕾,等.血管内皮生长因子对生物衍生骨复合骨髓基质细胞体内成骨的影响.中国临床康复,2003,7(11)：1622.
[13]Abramovitoh R, Neeman M, Reich R, et al. Intercellular communication between vascular smooth muscle and endothelial cells mediated by heparin-binding epidermal growth factor like growth factor and vascular endothelial growth factor. FEBS Letters 1999,425(4):441.
[14]Shifren TL, Doldi N, Ferrara N, et al. In the human fetus vascular endothelial growth factor is expressed in epithelisl cells and myocytes, but not vascular endothelium:Implications form odreofaction. J Chon Endocrinol Melab 1998:79(7):316.
[15]Steinbreeh DS, Mehrara BJ, Saodechp B, et al. Hypoxia regulates VEGF expression and cellular proliferation by osteoblasts in vitro. Plost ReConstr Surg 1999:104(3):738.
[16]芮钢,王者生,刘戈飞,等.人端粒酶反转录酶稳定转染成骨细胞构建组织工程骨的实验研究.中国临床康复.2004,8(4)：2613.
[17]Hendrix MJ,Seftor EA,Kirschmann DA,et al.Molecular biology of breast cancer metastasis molecular expression of vascular markers by aggressive breast cancer cells.Breasr Cancer Res.2000,2:417～422.
[18]Ruck P,Xiao JC, Kajserling E. Immunoreactivity of sinusoids in hepatcellular carcinoma. Arch Pathol Lab Med.1995,119:173～178.
[19]Reilly MS, Boehm T, Shing Y, et al. Endostatin:an endogenous inhibitor of angiogenesis and tumor growth. Cell.1997,88(2):277～285.
[20]宋继谒.病理学.科学出版社.1999：148.
[21]Kanwar YS.Biophysiology of glomerular filtration and proteinuia.Lab Invest.1984,51:7～ 21.
[22]A.A.C. Reed, C.J. Joyner, H.C. Browiilow, et al. Human atrophic fracture non-unions are not avascular. Journal of Orthopaedic Research.2002,(20):593～599.
[23]谢传高,王兴鹏,徐选福等.选择性环氧合酶-2抑制剂Celebrex对胰腺癌新生血管生成的影响.基础医学与临床.2004,24(6)：660-663.
[24]李胜棉,姚树坤.不同肝病变组织中cD34、cD31、Ki267的表达及意义.中国组织化学与细胞化学杂志,2004,13：70～75.
[25]Ruck P,Xiao JC, Kajserling E. Immunoreactivity of sinusoids in hepatcellular carcinoma. Arch Pathol Lab Med,1995,119:173～178.
[26]Horak ER, Leek R, Klenk N, et al. Angiogenesis, assessed by platelet/endothelial cell adhesion molecule antibodies, as indicator of node metastases and survival in breast cancer. Lancel,1992,340:1120-1124.
[27]Wu xx, Gordon RE, Robert W, et al. Morphological relationship ol von willebrand factor, typeVI collagen and fibrillin in human vascular subendothelium. Am J Pathol,1996,149: 283～291.
[28]张永刚,卢世璧,王继芳.骨引导与骨诱导.中华创伤杂志,1990,6：332～333.
[29]Humphries MJ. The molecular basis and specificity of intergrin-ligand interactions. J Cell Sci,1990,97:585-592.
[30]HILL AG, FINN P, SCHROEDER D. Postoperative fatigue after laparoscopic surgery. Aust N Z J Surg,1993,63(12):946～951.
[31]Mizuno M, Fujisawa R, Kuboki Y. Type I collagen-induced osteoblastic difrerentiation Of bone-marrow cell mediated by collagena2β1 interin interaction. Cell Physiol,2000,184: 207～13.
[32]Mizuno M, Kuboki Y. Osteoblast-related gene expression of bone marrow cells during the osteoblastic difrerentiation induced by type I collagen. J Biochem(Tokyo),2001,129:133～ 138.
[33]Suzawa M, Tarura Y, Fukumoto S, et al. Stimulation of Smadl transcriptional activity by Ras-extracellular signal-regulated kinase pathway:a possible mechanism for collagen-depending osteoblastic differentiation. J Bone Miner Res,2002,17:240～248.
[34]雪原,王沛,齐清会,等.淫羊藿甙对成骨细胞smad4 mRNA作用的试验研究.中华骨科杂志,2005,25：119～123.
[35]Seftor RE. Role of the [33 integrin subunit in human primary melanoma progression. Am J Path,1998,153(5):1347～1351.
[36]Lin EC, Carron CP,Meyer DM, et al. A series of function blocking antibodies against the alpha V beta 3 integrin bind allosteric to the ligand binding site and induce ligand dissociation. Cell Adhes Commun,1998,6(6):451～464.
[37]Drake CJ, Cheresh DA, Little CD. An antagonist of integrin alpha V beta 3 prevent maturation of blood vessels during embryonic neovascularization. J Cell Sci,1995,108: 2655～2661.
[1]张俐,刘献佯.中医药对骨折治疗的研究.中国中医骨伤科杂志.1997.5(6)：51.
[2]侯晓峰,刘景生.中药促进骨折愈合机理.中国中医骨伤科杂志.2003,11(1)：61～63.
[3]申海明.近30年来云南白药应用状况[J].云南医药杂志,1986,7(4)：46.
[4]朱大忠等.云南白药的药理及临床研究进展.中西医结合杂志.1987：7(6)：377～379.
[5]胥沙订,葛宝丰,徐印坎.实用骨科学[M].北京：人民军医出版社,2003.2.
[6]王亦璁.骨与关节损伤[M].北京：人民卫生出版社,2001.149～154.
[7]Brighton CT. Principle of fracture healing factors in fracture healing. Clin Orthop.1992,277～ 297.
[8]Ross R, Raines EW. The biology of platelet derived growth factor. Cell,1986,46:115.
[9]Sporn MB, Roberts AB. Some recent advances in the chemstry of transforming growth factor beta. Cell Biol,1987,105:1039.
[10]Sprugel KH, Mcpherson J. Effects of growth factors in vivo. Am J Pathol,1987,129:601.
[11]杜俊杰,罗卓荆,胡蕴玉,等.rhBMP-2在体内诱导成骨中Ⅰ、Ⅱ型胶原及碱性磷酸酶的表达[J].第四军医大学学报,2001,22(11)：981～983.
[12]胡德志,付刚.骨形态发生蛋白2(BMP-2)的研究进展[J].中医正骨,1998,10(1)：47～49.
[13]Pathis, Rntenberg JB, Johnson RL, et al. Interaction of Ihh and BMP/Noggin signaling during cartilage differentiation. Dev Biol,1999,209(2):239～253.
[14]苏佳灿,王家林,张春才.成骨蛋白-7促进骨折愈合研究进展[J].中国矫形外科杂志,2001,8(6)：599～601.
[15]Shifren TL, Doldi N, Fermra N, et al. In the human fetus, vascular endothelial growth factor is expressed in epithelial cells and myocytes, but not vascular endothelium:implications for mode of action. J Clin Endocrinol Melab,1994,79(1):316～322.
[16]Tanaka T, Taniguchl Y, Gotoh K, et al. Morphological study of recombinant human transforming growth factor betal-induced intramembranous ossification in neonatal rat pailet bone. Bone,1993,14(2):117-123.
[17]Koyama H. Analysis of genes implicated in osteogenic signaling by osteogenic protein-1 (OP-1) using differential display method. Kikubyo Gakkai Zasshi,1997,64(2):429-431.
[18]Rundle CH, Miyakoshi N, Ramirez E, et al. Expression of the fibroblast growth factor receptor genes in fracture repair. J Clin Orthop,2002, (403):253-263.
[19]刘润田.中药接骨丹对骨折愈合作用的研究[J],天津医药,1962,4(8)：457.
[20]李小生,唐杨,王茂泓.“肾主骨”与肾性骨病理论探讨[J].江西中医药,2006,37(279)：12～-13.
[21]阮朝阳,祁开泽,中药促进骨折愈合作用机制的实验研究进展.湖南中医药导报,2003, 9(2)：48～49.
[22]胡美珠.丹参注射液对家兔桡骨骨折以后肢体血容量的影响[J].中华外科杂志,1979；17(1)：33.
[23]侯筱魁.外敷中药对家兔桡骨骨折愈合的影响[J].中国中医骨伤科杂志,1992,8(2)：10.
[24]贵阳中医学院骨折科研协作组.中药“接骨Ⅱ号”促进骨折愈合作用的实验观察[J].中华医学杂志,1978,11：670.
[25]曲克服等.理气活血剂在骨折愈合过程生化和生物力学观察[J].上海中医药杂志,1982,5：42.
[26]吴汝舟等.微量元素对家兔骨折愈合影响的实验研究[J].中华骨科杂志,1992,12(4)：282.
[27]王智光等.自然铜对家兔骨折后骨痂中的胶原、钙、磷等的影响[J].中华外科杂志,1986,6(4)：305.
[28]张菊英.丹参注射液对骨折愈合中钙沉积的影响[J].中西医结合杂志,1984,(9)：536.
[29]胡美珠.丹参注射液对家兔桡骨骨折后全身血容量的影响[J].中华外科杂志,1979,17(1)：33.
[30]夏志道.壮骨素促进骨折愈合的实验研究[J].中国骨伤,1994,7(1)：7.
[31]孙炳烈.平乐内服接骨丹对骨折愈合作用的实验研究[J].中西医结合杂志,1995,7(9)：544.
[32]周秋丽.鹿茸总多肽促进软骨及成骨样细胞增殖及骨折愈合[J].中国药理学报,1999,20(3)：279～282.
[33]王宝泉.生肌象皮膏在感染开放性骨折中的实验研究[J].中医杂志,1981,1：25.
[34]杨淮纭.中草药常绿油麻藤种子提取左旋多巴促进骨折愈合的临床及实验研究[J].中西医结合杂志.1985,5：7.
[35]林燕萍.锻狗骨对实验性骨折愈合作用的垂体GH,TSH细胞免疫化学研究[J].中国骨伤1993,6(5)：5.
[36]刘献样等.麦饭石促进骨折愈合的实验研究[J].中国中医骨伤科杂志.1995,3(2)：5.
[37]秦军志等.丹参对大鼠胫骨骨折早期愈合过程中血清、骨痂及骨组织中钙、锌、铜的影响[J].中国中西医结合杂志1992；12(6)：354.
[38]马克昌.益气化瘀汤对实验性骨折的影响[J].中医正骨1995,7(1)：3.
[39]魏玉玲.伤科接骨片促进骨折愈合作用的实验研究[J].中医正骨,1996；8(4)：3.
[40]刘艺,顾国奎,刘克泉,等.海螵蛸接骨动物实验的组织学研究[J].中国中医骨伤科杂志,1995,3(5)：6～8.
[41]杨庆秋,王少飞,胡侦明等.云南白药促进骨缺损修复及引导性骨再生的实验研究[J].中国矫形外科杂志,2002,(08)：65～66.
[42]高崇昆主编.云南白药研究与应用[M].昆明：云南科技出版社,2002：39～40.
[43]张慧,班立丽,杨兵兵等.云南白药含药血清对大鼠骨髓间充质干细胞增殖和成骨分化的影响.中国科技论文在线.1～10.
[44]许建阳,王发强,郑维发等.金铁锁对实验性RA小鼠痛阈及血清NO/NOS含量的影响[J].中医药学刊,2004,22(1)：82～84.
[45]叶剑锋,严伟民,甘卓慧等.云南白药对大鼠血小板聚集及膜糖蛋白表达的影响[J].中国现代应用药学杂志,2004,21(2)：100～103.
[46]张昌钦,邓应忠,沈志祥.云南白药灌肠治疗下消化道出血疗效观察[J].中华全科医师杂志,2005,4(2)：126.
[47]王成坤,蔡家骏,黄洋等.云南白药修复犬牙实验性髓室底穿孔的组织学研究[J].中国中西医结合杂志,1994,14(6)：357.
[48]季申,周坛树,张锦哲.中药重楼和云南白药中抗肿瘤细胞毒活性物质Gracillin的测定[J].中国中西医结合杂志,2001,23(3)：212～215.
[49]施莱.中药加速骨折愈合机理的研究.长春中医药大学学报.2010,26(8)：600～602.
[2]潘振球等.单味云南白药外敷的应用概况.中国临床医生.2001；29(7)：49.
[3]朱大忠等.云南白药的药理及临床研究进展.中西医结合杂志.1987：7(6)：377～379.
[4]蔡锡麟等.云南白药的药理研究.中成药研究.1982：(8)： 37.
[5]孔令义,杨智,闵知大.对中药现代化中有效成分研究的思考.中草药,1998,29(5)：354～355.
[6]刘建立.中药复方疗效的物质基础及其化学成分研究方法.中成药,1992,14(10)：49.
[7]王永炎,赵宜军.从大黄复方的运用浅谈中药复方研究的方法与思路.中国中药杂志,2000,25(3)：131～132.
[8]何祥久,邱峰,姚新生.中药复方研究现状和思路.化学进展,2001,13(6)：481～484.
[9]罗国安,王义明.中药复方物质基础和药效相关性研究,世界学技术-中药现代化,1999,1：11～15.
[10]武孔云,梁光义,靳风云,等.中药复方药效物质基础研究的思路及方法.世界科学技术-中医药现代化,2003,5(6)：13～15.
[11]孙明瑜,谢鸣.试论证候与复方的研究思路.中国医药学报,2002,17(4)：196～198.
[12]姚新生,胡柯.中药复方的现代化研究.化学进展。1999,11(2)：192～196.
[13]程永现,赵淑敏.中药复方研究中的几个关键性问题-中医现代化思路探索.中国医药学报,2001,16(1)：63-65.
[14]徐东铭,徐雅红.中药复方药效活性成分研究思路初探.中国医药学报,2001,16(6)：52-54.
[15]刘良.中药复方基础研究的思路与方法探讨.中国实验方剂学杂志,2000,6(4)：1～4.
[16]方文贤.中药新药研究开发与思路创新的再思考.中国医药学报,2003,18(6)：365～369.
[17]李澎涛,乔延江,王永炎,等.对中药复方研究的思考.北京中医药大学学报,1999,22(5)：15～18.
[1]王和鸣,王力,李楠.巴戟天对骨髓基质细胞向成骨细胞分化影响的实验研究.福建中医学院学报.2004,14(3)：16～20.
[2]王银喜,王金熙,徐智敏,等.局部植入接骨中药提取物促进骨愈合的实验研究.中国骨伤,2003；16(6)：346～349.
[3]王斌,罗毅文,胡年宏.骨康方对大鼠MSCs体外向成骨细胞分化和ALP的影响.中国中医骨伤科杂志.2007,15(12)：5～9.
[4]韩立民,刘波,徐彭.淫羊藿对成骨细胞增殖的血清药理学研究.中医药学刊,2003,21(5)：678～680.
[5]祝晓玲,祝彼得.黄芪体外作用对贫血小鼠骨髓基质细胞分泌SCF的影响.细胞与分子免疫学杂志,2002,18(4)：396.
[6]Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precurors in normal and irradiated mouse hematopoietic organs. Exp Hematol,1976,4(5):267～74.
[7]Caplan AL. The mesengenic process. Clin Plast Surg,1994,21:429～435.
[8]Pitlenger MF, Mackay AM, Beck SC et al. Multilineage potential of adult human mesenchymal stem cells. Science,1999,284:143～147.
[9]Deans RJ, Moseley AB. Mesenchymal stem cells:Biology and potential clinical uses. Exp Hematol,2000,28:875～884.
[10]Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potennial of adult human mesenchymal stem cells. Science,1999,284:143～147.
[11]Nathanson MA. Bone matrix-directed chondrogenesis of muscle in vitro. J Clin Orthop, 1985,200:142～158.
[12]Nakahara H, Dennis J E, Bruder SP, et al. In vitro differentiation of bone and hypertrophic cartilage from periosteal-derived cells. Exp Cell Res,1991,195:492～503.
[13]Campagnoli C, Roberts IA, Kumar S, et al. Identification of mesenchymal stem/progenitor cells in human first-trimester fetal blood, liver, and bone marrow. Blood,2001,98:2396～ 2402.
[14]Chen TL, Shen WJ, Kraemer FB. Human BMP-7/0P-1 induces the growth and differentiation Of adipocytes and osteoblasts in bone marrow stromal cell cultures. Cell Biochem.2001, 82(2):187.
[15]支会英,李恩.补肾方药对去卵巢骨质疏松的骨组织ER表达的影响.中国中医基础医 学杂志,2001,7(4)：41～44.
[16]Rhee ST, EI-Bassiony L, Buchman SR, et al. Extracellular signal-related kinase and bone morphogenetic protein expression during distraction osteogenesis of the manndible:in vivo evidence of a mechanotransduction mechanism for differentiation and osteogenesis by mesenchymal precursor cells. Plast Reconstr Surg,2006,117(7):2243.
[17]郝瑞锋.申长虹.骨髓问充质干细胞向成骨细胞诱导的研究现状.天津医科大学学报。2007,13(3)：129～131.
[18]RINGE J, KAPS C, BURMESTER G R, et al. Stem cells for re—generative medicine: advances in the engineering of tissues and organs(Review). Naturwissenschaften,2002, 89(8):38-351.
[19]CLARKE D, FRISEN J. Differentiation potential of adult stem cells. Curr Opin Genet.2001, 11(5):575-80.
[20]Ross R, Raines EW, et al. The biology of platelet derived growth factor. Cell,1986,46:115.
[21]Robey PG, Yong MF, et al. Osteo-blasts synthesize and respond to transforming growth factor beta in vitro. J Cell Biol,1987,105:457.
[22]Andrew JW, Hoyland J, et al. Demonstraction of BMP2 mRNA expression in human fracture callus. J Bone Joint Surg,1993,75:(supp3),88.
[23]Cook-SD, Salkeld-SL, Brinker-MR, et al. Use of an osteoinductive biomaterial (rhOP-1) in healing large segmental bone defects. J Orthop-Trauma.1998,8; 12(6):407～412.
[24]DAVID C, REINER C, CARLA M, et al. Rapid expansion of recycling stem cells in cultures of plastic adherent cells from human bone marrow. Pronc Natl Acad Sci,2000,97(7):3213～ 3218.
[25]Brian Jonstone, Tomas M. In vitro Chondrogenesis of bone marrow-derived mesenchymal Progenitor cells. Experiment Cell Resarch,1998,238:265～271.
[26]夏万尧,商庆新,崔磊,等.骨髓间充质干细胞向软骨细胞表型定向诱导分化的实验研究[J].中华整形外科杂志,2002,18：12～14.
[27]Noble BS, Dean V, loueridge N. Dextran sulfate promotes the rapid aggregation of porcine bone-marrow stromal cells. Bone,1995,1(4):375～382.
[28]杨志明.组织工程基础与临床成都[M].成都：四川科学技术出版社,2000,37～57.
[29]Long MW, Robinson JA, A shcraft EA, et al. Regulation of human bone marrow-derived osteoprogenitor cells by osteogenic growth factors. J Clin Invest,1995,95(2):881.
[30]王蓓,汪维伟.问充质干细胞的研究进展[J].国外医学：外科学分册,2005,32(6)： 459～462.
[31]Turksen K, Aubin J E. Positive and negative immunoselection for enrichment of two classes of osteoprogenitor cells. J Cell Biol,1991,114:373～384.
[32]Maniatopoulos C, Sodek J, MelcherAH. Bone formation in vitro by stromal cells obtained from bonemarrow of young adult rats. Cell Tissue Res,1998,254:317.
[33]McQillan DJ, Richardson MD, Bateman JF. Matrix deposition by a calcifying human osteogenic sarcoma cell line (SAOS22). Bone,1995,16:415～426.
[34]Otsuka E, Yamaguchi A, Shigehisa H, et al. Characterizations of osteoblastic differentiation of stromal cell line ST2 that is induced by ascorbicacid. Am J Physiol,1999,277:132～138.
[35]Masaki I, Takenbu K, Shuichi A, et al. Bone morphogenetic protein-12 and-13 inhibit terminal differentiation of myoblasts, but do not induce their differentiation into osteoblast. Biochem Biophys Res Commun,1996,222:317～322.
[36]Kim KJ, Itoh T, Kotake S. Effects of recombinant human bone morphogenetic protein-2 on human bone marrow cells culture with various biomaterial. Biomed Mater Res,1997,35: 279～285.
[37]Gronthos S, Zannetion AC, Hay SJ, et al. Molecular and celluer characterization of highly purified stromal stem cells derived from human bone marrow. J Cell Sci,2003,116:1827～ 1835.
[38]Tsuda H, Wada T, Ito Y, et al. Efficient BMP2 gene transfer and bone formation of mesenchymal stem cells by a fiber-mutant adenoviral vector. J Mol Ther,2003,7:354～365.
[39]Rickard Dj, Sullivan TA, Shenker BJ, et al. Induction of rapid osteoblast differentiation in rat bone marrow stromal cell culture by dexamethasone and BMP-2. J Dev Biol,1994,161: 218～228.
[40]杨丕斌,胡侦明,劳汉昌.雌激素对骨折愈合过程中骨形态发生蛋白-4 mRNA表达的影响.中国矫形外科杂志,2003,5：620～621.
[41]Zhenming Hu, Sean A.F. Peel, George K. B. Sander, Cameron M. L. Clokie. The osteoinductive activity of bone morphogenetic protein (BMP) purified by repeated extracts of bovine bone. Growth Factors,2004,2:1～5.
[42]Rosier RN O, Keefe RJ, Hicks DG. The potential role of transforming growth factor beta in fracture healing. Clin Orthop.1998, (355 Suppl):294～300.
[1]柴本甫.骨折模型[J].中华外科杂志.1962.1(5)：299-301.
[2]李人杰,柳用墨,王琮武.旋磁场促进骨折愈合的初步实验研究.中华骨科杂志,1998,8(6)：445～447.
[3]Kraan MC, Smith MD, Weedon H et al. Measurement of cytokine and adhesion molecule expression in synovial tissue by digital image analysis. Ann Rheum Dis,2001;60; 296～298.
[4]吕翔.病理图像定量分析及其测量误差的控制.中国体视学与图像分析.2002,7(1)：58～62.
[5]Frost HM. Magovem CJ, Mack CA, et al. The biology of fracture healing. an overview for clinicians. Clin Orthop 1992:248(11):294.
[6]Arobson J. Cassnano C. Rowhim M, et al. Experimental and clinical expcrience with distraction osteogenesis. ClEfipalate Craniofac 1995:31(6):473.
[7]Eppley BL, Connolly DT, Winkelmann T, et al. Free bone graft reconstruction of irradiated facial tissue:Experimental effects of basic fibroblast growth factor stimulation. Plast Reconstr Surg 1996:88(1):1.
[8]Trueta P, Sacehi N, Lei J, et al. Single-step method of RNA is olation by acid guanidinium thiocyanate—phen01. chloroform extraction. Anal Biochem 1997; 162(1):156.
[9]Chuanyong Lu, Erik Hansen, Anna Sapozhnikova,et al.Effect of Age on Vascularization during Fracture Repair.J Orthop Res.2008,26:1384-1389.
[10]Ru Li, Duncan J. Stewart, Herbert P,et al.Effect of Cell-Based VEGF Gene Therapy on Healing of a Segmental Bone Defect.J Orthop Res.2009,27:8-14.
[11]FelTara N. Davis Smyth T. Vincenti V, et al. The biology of vascular endothelial growth factor. Endocr Res 1997:18(8):4.
[12]刘杰,孙正义,曹蕾,等.血管内皮生长因子对生物衍生骨复合骨髓基质细胞体内成骨的影响.中国临床康复,2003,7(11)：1622.
[13]Abramovitoh R, Neeman M, Reich R, et al. Intercellular communication between vascular smooth muscle and endothelial cells mediated by heparin-binding epidermal growth factor like growth factor and vascular endothelial growth factor. FEBS Letters 1999,425(4):441.
[14]Shifren TL, Doldi N, Ferrara N, et al. In the human fetus vascular endothelial growth factor is expressed in epithelisl cells and myocytes, but not vascular endothelium:Implications form odreofaction. J Chon Endocrinol Melab 1998:79(7):316.
[15]Steinbreeh DS, Mehrara BJ, Saodechp B, et al. Hypoxia regulates VEGF expression and cellular proliferation by osteoblasts in vitro. Plost ReConstr Surg 1999:104(3):738.
[16]芮钢,王者生,刘戈飞,等.人端粒酶反转录酶稳定转染成骨细胞构建组织工程骨的实验研究.中国临床康复.2004,8(4)：2613.
[17]Hendrix MJ,Seftor EA,Kirschmann DA,et al.Molecular biology of breast cancer metastasis molecular expression of vascular markers by aggressive breast cancer cells.Breasr Cancer Res.2000,2:417～422.
[18]Ruck P,Xiao JC, Kajserling E. Immunoreactivity of sinusoids in hepatcellular carcinoma. Arch Pathol Lab Med.1995,119:173～178.
[19]Reilly MS, Boehm T, Shing Y, et al. Endostatin:an endogenous inhibitor of angiogenesis and tumor growth. Cell.1997,88(2):277～285.
[20]宋继谒.病理学.科学出版社.1999：148.
[21]Kanwar YS.Biophysiology of glomerular filtration and proteinuia.Lab Invest.1984,51:7～ 21.
[22]A.A.C. Reed, C.J. Joyner, H.C. Browiilow, et al. Human atrophic fracture non-unions are not avascular. Journal of Orthopaedic Research.2002,(20):593～599.
[23]谢传高,王兴鹏,徐选福等.选择性环氧合酶-2抑制剂Celebrex对胰腺癌新生血管生成的影响.基础医学与临床.2004,24(6)：660-663.
[24]李胜棉,姚树坤.不同肝病变组织中cD34、cD31、Ki267的表达及意义.中国组织化学与细胞化学杂志,2004,13：70～75.
[25]Ruck P,Xiao JC, Kajserling E. Immunoreactivity of sinusoids in hepatcellular carcinoma. Arch Pathol Lab Med,1995,119:173～178.
[26]Horak ER, Leek R, Klenk N, et al. Angiogenesis, assessed by platelet/endothelial cell adhesion molecule antibodies, as indicator of node metastases and survival in breast cancer. Lancel,1992,340:1120-1124.
[27]Wu xx, Gordon RE, Robert W, et al. Morphological relationship ol von willebrand factor, typeVI collagen and fibrillin in human vascular subendothelium. Am J Pathol,1996,149: 283～291.
[28]张永刚,卢世璧,王继芳.骨引导与骨诱导.中华创伤杂志,1990,6：332～333.
[29]Humphries MJ. The molecular basis and specificity of intergrin-ligand interactions. J Cell Sci,1990,97:585-592.
[30]HILL AG, FINN P, SCHROEDER D. Postoperative fatigue after laparoscopic surgery. Aust N Z J Surg,1993,63(12):946～951.
[31]Mizuno M, Fujisawa R, Kuboki Y. Type I collagen-induced osteoblastic difrerentiation Of bone-marrow cell mediated by collagena2β1 interin interaction. Cell Physiol,2000,184: 207～13.
[32]Mizuno M, Kuboki Y. Osteoblast-related gene expression of bone marrow cells during the osteoblastic difrerentiation induced by type I collagen. J Biochem(Tokyo),2001,129:133～ 138.
[33]Suzawa M, Tarura Y, Fukumoto S, et al. Stimulation of Smadl transcriptional activity by Ras-extracellular signal-regulated kinase pathway:a possible mechanism for collagen-depending osteoblastic differentiation. J Bone Miner Res,2002,17:240～248.
[34]雪原,王沛,齐清会,等.淫羊藿甙对成骨细胞smad4 mRNA作用的试验研究.中华骨科杂志,2005,25：119～123.
[35]Seftor RE. Role of the [33 integrin subunit in human primary melanoma progression. Am J Path,1998,153(5):1347～1351.
[36]Lin EC, Carron CP,Meyer DM, et al. A series of function blocking antibodies against the alpha V beta 3 integrin bind allosteric to the ligand binding site and induce ligand dissociation. Cell Adhes Commun,1998,6(6):451～464.
[37]Drake CJ, Cheresh DA, Little CD. An antagonist of integrin alpha V beta 3 prevent maturation of blood vessels during embryonic neovascularization. J Cell Sci,1995,108: 2655～2661.
[1]张俐,刘献佯.中医药对骨折治疗的研究.中国中医骨伤科杂志.1997.5(6)：51.
[2]侯晓峰,刘景生.中药促进骨折愈合机理.中国中医骨伤科杂志.2003,11(1)：61～63.
[3]申海明.近30年来云南白药应用状况[J].云南医药杂志,1986,7(4)：46.
[4]朱大忠等.云南白药的药理及临床研究进展.中西医结合杂志.1987：7(6)：377～379.
[5]胥沙订,葛宝丰,徐印坎.实用骨科学[M].北京：人民军医出版社,2003.2.
[6]王亦璁.骨与关节损伤[M].北京：人民卫生出版社,2001.149～154.
[7]Brighton CT. Principle of fracture healing factors in fracture healing. Clin Orthop.1992,277～ 297.
[8]Ross R, Raines EW. The biology of platelet derived growth factor. Cell,1986,46:115.
[9]Sporn MB, Roberts AB. Some recent advances in the chemstry of transforming growth factor beta. Cell Biol,1987,105:1039.
[10]Sprugel KH, Mcpherson J. Effects of growth factors in vivo. Am J Pathol,1987,129:601.
[11]杜俊杰,罗卓荆,胡蕴玉,等.rhBMP-2在体内诱导成骨中Ⅰ、Ⅱ型胶原及碱性磷酸酶的表达[J].第四军医大学学报,2001,22(11)：981～983.
[12]胡德志,付刚.骨形态发生蛋白2(BMP-2)的研究进展[J].中医正骨,1998,10(1)：47～49.
[13]Pathis, Rntenberg JB, Johnson RL, et al. Interaction of Ihh and BMP/Noggin signaling during cartilage differentiation. Dev Biol,1999,209(2):239～253.
[14]苏佳灿,王家林,张春才.成骨蛋白-7促进骨折愈合研究进展[J].中国矫形外科杂志,2001,8(6)：599～601.
[15]Shifren TL, Doldi N, Fermra N, et al. In the human fetus, vascular endothelial growth factor is expressed in epithelial cells and myocytes, but not vascular endothelium:implications for mode of action. J Clin Endocrinol Melab,1994,79(1):316～322.
[16]Tanaka T, Taniguchl Y, Gotoh K, et al. Morphological study of recombinant human transforming growth factor betal-induced intramembranous ossification in neonatal rat pailet bone. Bone,1993,14(2):117-123.
[17]Koyama H. Analysis of genes implicated in osteogenic signaling by osteogenic protein-1 (OP-1) using differential display method. Kikubyo Gakkai Zasshi,1997,64(2):429-431.
[18]Rundle CH, Miyakoshi N, Ramirez E, et al. Expression of the fibroblast growth factor receptor genes in fracture repair. J Clin Orthop,2002, (403):253-263.
[19]刘润田.中药接骨丹对骨折愈合作用的研究[J],天津医药,1962,4(8)：457.
[20]李小生,唐杨,王茂泓.“肾主骨”与肾性骨病理论探讨[J].江西中医药,2006,37(279)：12～-13.
[21]阮朝阳,祁开泽,中药促进骨折愈合作用机制的实验研究进展.湖南中医药导报,2003, 9(2)：48～49.
[22]胡美珠.丹参注射液对家兔桡骨骨折以后肢体血容量的影响[J].中华外科杂志,1979；17(1)：33.
[23]侯筱魁.外敷中药对家兔桡骨骨折愈合的影响[J].中国中医骨伤科杂志,1992,8(2)：10.
[24]贵阳中医学院骨折科研协作组.中药“接骨Ⅱ号”促进骨折愈合作用的实验观察[J].中华医学杂志,1978,11：670.
[25]曲克服等.理气活血剂在骨折愈合过程生化和生物力学观察[J].上海中医药杂志,1982,5：42.
[26]吴汝舟等.微量元素对家兔骨折愈合影响的实验研究[J].中华骨科杂志,1992,12(4)：282.
[27]王智光等.自然铜对家兔骨折后骨痂中的胶原、钙、磷等的影响[J].中华外科杂志,1986,6(4)：305.
[28]张菊英.丹参注射液对骨折愈合中钙沉积的影响[J].中西医结合杂志,1984,(9)：536.
[29]胡美珠.丹参注射液对家兔桡骨骨折后全身血容量的影响[J].中华外科杂志,1979,17(1)：33.
[30]夏志道.壮骨素促进骨折愈合的实验研究[J].中国骨伤,1994,7(1)：7.
[31]孙炳烈.平乐内服接骨丹对骨折愈合作用的实验研究[J].中西医结合杂志,1995,7(9)：544.
[32]周秋丽.鹿茸总多肽促进软骨及成骨样细胞增殖及骨折愈合[J].中国药理学报,1999,20(3)：279～282.
[33]王宝泉.生肌象皮膏在感染开放性骨折中的实验研究[J].中医杂志,1981,1：25.
[34]杨淮纭.中草药常绿油麻藤种子提取左旋多巴促进骨折愈合的临床及实验研究[J].中西医结合杂志.1985,5：7.
[35]林燕萍.锻狗骨对实验性骨折愈合作用的垂体GH,TSH细胞免疫化学研究[J].中国骨伤1993,6(5)：5.
[36]刘献样等.麦饭石促进骨折愈合的实验研究[J].中国中医骨伤科杂志.1995,3(2)：5.
[37]秦军志等.丹参对大鼠胫骨骨折早期愈合过程中血清、骨痂及骨组织中钙、锌、铜的影响[J].中国中西医结合杂志1992；12(6)：354.
[38]马克昌.益气化瘀汤对实验性骨折的影响[J].中医正骨1995,7(1)：3.
[39]魏玉玲.伤科接骨片促进骨折愈合作用的实验研究[J].中医正骨,1996；8(4)：3.
[40]刘艺,顾国奎,刘克泉,等.海螵蛸接骨动物实验的组织学研究[J].中国中医骨伤科杂志,1995,3(5)：6～8.
[41]杨庆秋,王少飞,胡侦明等.云南白药促进骨缺损修复及引导性骨再生的实验研究[J].中国矫形外科杂志,2002,(08)：65～66.
[42]高崇昆主编.云南白药研究与应用[M].昆明：云南科技出版社,2002：39～40.
[43]张慧,班立丽,杨兵兵等.云南白药含药血清对大鼠骨髓间充质干细胞增殖和成骨分化的影响.中国科技论文在线.1～10.
[44]许建阳,王发强,郑维发等.金铁锁对实验性RA小鼠痛阈及血清NO/NOS含量的影响[J].中医药学刊,2004,22(1)：82～84.
[45]叶剑锋,严伟民,甘卓慧等.云南白药对大鼠血小板聚集及膜糖蛋白表达的影响[J].中国现代应用药学杂志,2004,21(2)：100～103.
[46]张昌钦,邓应忠,沈志祥.云南白药灌肠治疗下消化道出血疗效观察[J].中华全科医师杂志,2005,4(2)：126.
[47]王成坤,蔡家骏,黄洋等.云南白药修复犬牙实验性髓室底穿孔的组织学研究[J].中国中西医结合杂志,1994,14(6)：357.
[48]季申,周坛树,张锦哲.中药重楼和云南白药中抗肿瘤细胞毒活性物质Gracillin的测定[J].中国中西医结合杂志,2001,23(3)：212～215.
[49]施莱.中药加速骨折愈合机理的研究.长春中医药大学学报.2010,26(8)：600～602.