一种智能化电穿孔仪的实现及毫米波生物效应的研究
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摘要
近年来，人们在电磁场产生的生物学效应及应用的研究越来越广泛，这其中包括对连续电磁波的生物热效应和非热生物效应，以及对瞬态电磁场与生物体的相互作用影响等方面。
电穿孔效应是指在适当高压脉冲电场作用下，细胞或组织间起相对隔离作用的“屏障”内快速形成液态通道的现象，是电场与具有不同介电常数而且易变形的物质相互作用的结果。本文从孔的形成、阈值跨膜电压、膜损伤、分子运输、组织穿孔、膜恢复和细胞死亡等方面论述了电穿孔效应的产生机理，并对电穿孔效应的应用进行了总结，在此基础上，结合电穿孔生物效应实验研究和应用特点，设计制作了一台智能化电穿孔仪，该仪器具有矩形脉冲和电容放电脉冲两种波形输出，并且脉冲幅值、脉冲宽度、重复频率可调，文中对它的工作原理、系统结构、性能参数作了简要说明。
毫米波(MMW)生物效应也是人们研究的一个重要方面。本文在介绍了几种典型的毫米波生物效应，并对其应用以及目前在毫米波生物效应机理研究方面的各种理论模型作了概括后，对毫米波作用人舌癌细胞产生的效应进行了较系统的研究，主要集中在细胞对毫米波吸收的频率和强度的选择性问题上，最后对实验结果进行了分析讨论。
In recent years, the biological and medical effects of electromagnetic field has been studied widely including the thermal and non-thermal effects of continuous electromagnetic wave and the effects of transient electromagnetic field.
The essentially universal biophysical phenomenon of "electroporation" occurs if an appropriate pulse field is applied. Electroporation is believed to be the rapid creation of aqueous pathways through lipid-containing barriers in cells and tissue. The driving force is the physical interaction of electric fields with different dielectric constants. This thesis introduced the phenomenon of electroporation and gave some discussions on the mechanism of electroporation in various aspects such as pore formation, membrane damage, tissue electroporation, membrane recovery and cell death and so on. This thesis designed a intellectualized electroporation instrument which can output rectangular pulse and capacitor discharge pulse. The voltage output, pulsewidth and repetition can be variable. The principle, hardware and software system and specifications were introduced in this paper.
The research of biological medical effects of millimeter wave (MMW) has been of compelling interest. This thesis reviewed the new effects of MMW and some representative opinions about the mechanism of MMW effects. The inducing effect of MMW on tongue cancer cell has been researched in this paper. According to the research result, this thesis gave a discussion on the problem of frequency and power of MMW.
电穿孔效应是指在适当高压脉冲电场作用下，细胞或组织间起相对隔离作用的“屏障”内快速形成液态通道的现象，是电场与具有不同介电常数而且易变形的物质相互作用的结果。本文从孔的形成、阈值跨膜电压、膜损伤、分子运输、组织穿孔、膜恢复和细胞死亡等方面论述了电穿孔效应的产生机理，并对电穿孔效应的应用进行了总结，在此基础上，结合电穿孔生物效应实验研究和应用特点，设计制作了一台智能化电穿孔仪，该仪器具有矩形脉冲和电容放电脉冲两种波形输出，并且脉冲幅值、脉冲宽度、重复频率可调，文中对它的工作原理、系统结构、性能参数作了简要说明。
毫米波(MMW)生物效应也是人们研究的一个重要方面。本文在介绍了几种典型的毫米波生物效应，并对其应用以及目前在毫米波生物效应机理研究方面的各种理论模型作了概括后，对毫米波作用人舌癌细胞产生的效应进行了较系统的研究，主要集中在细胞对毫米波吸收的频率和强度的选择性问题上，最后对实验结果进行了分析讨论。
In recent years, the biological and medical effects of electromagnetic field has been studied widely including the thermal and non-thermal effects of continuous electromagnetic wave and the effects of transient electromagnetic field.
The essentially universal biophysical phenomenon of "electroporation" occurs if an appropriate pulse field is applied. Electroporation is believed to be the rapid creation of aqueous pathways through lipid-containing barriers in cells and tissue. The driving force is the physical interaction of electric fields with different dielectric constants. This thesis introduced the phenomenon of electroporation and gave some discussions on the mechanism of electroporation in various aspects such as pore formation, membrane damage, tissue electroporation, membrane recovery and cell death and so on. This thesis designed a intellectualized electroporation instrument which can output rectangular pulse and capacitor discharge pulse. The voltage output, pulsewidth and repetition can be variable. The principle, hardware and software system and specifications were introduced in this paper.
The research of biological medical effects of millimeter wave (MMW) has been of compelling interest. This thesis reviewed the new effects of MMW and some representative opinions about the mechanism of MMW effects. The inducing effect of MMW on tongue cancer cell has been researched in this paper. According to the research result, this thesis gave a discussion on the problem of frequency and power of MMW.
引文
[1] J.C.Weaver. Electroporation in cells and tissues: A biophysical phenomenon due to electromagnetic fields. Radio Sci. , 1995, vol. 30, no. 1: 205-221
[2] Abidor. I. G, V.B.Arakelyan, L. V. Chernomordik and Y. A. Chizmadzhev. Electric breakdown of bilayer membranes,I. The main experimental facts and their qualitative discussionBioelectrochem.Bioenerg, 1979,6,37-52
[3] Barnett.A,and J. C. Weaver. Electroporation: A unified quantitative theory of reversible electrical breakdown and rupture. Bioelectrochem. Bioenerg. , 1991, 25, 163-182
[4] Weaver.J.C. Molecular basis for cell membrane electroporation. Proc.N. Y. Acad. Sci., 1993,720,141-152
[5] Chang.D.C, B. M.Chassy and J. A. Saunders. Guide to Electroporation and Electrofusion. Academic,San Diego,Calif,1992
[6] Lipowsky. R. The conformation of membranes. Nature, 1991, 349, 475-481
[7] Pastushenko. V.F. and Y. A. Chizmadzhev. Stabilization of conducting pores in BLM by electric current. 1982,1,43-52
[8] Powell. K. T. and J. C. Weaver. Transient aqueous pores in bilayer membranes: A statistical theory. Bioelectrochem.Bioelectroenerg. , 1986, 15, 211-227
[9] Sabelnikov.A.G. and E.S.Cymbalyuk. Clotting of donor DNA during electroporation of prokaryotes. Bioelectrochem. Bioenerg. 1990,24, 313-321
[10] Crowley.J.M. Electrical breakdown of bimolecular lipid membranes as an electromechanical instability. Biophys,1973,13,711-724
[11] Weaver.J.C. and R.A.Mintzer. Decreased bilayer stability due to transmembrane potentials. Phys. Lett. , 1981,86,57-59
[12] Suarg.I.P. The effects of external fields on the structure of lipid bilayers. J. Physiol. Paris. , 1981,77,1035-1042
[13] Weaver.J.C. and A.Barnett. Progress towards a theoretical model of electroporation mechanism: Membrane electrical behavior and molecular transport. In D. C. Chang, B. M. Chassy, J. A. Saunders, and A. E. Sowers, eds. Guide to electroporation and electrofusion . San Diego: Acdemic, 1992, 91-117
[14]Benz. R. and U. Zimmermann, Relaxation studies on cell membranes and lipid bilayers in high electric field range. Bioelectrochem. bioenerg.,1980,7, 723～739
[15]Zimmermann. U., P. Scheurich, G. Pilwat, and R. Benz Cells with manipulated function: New perspective for cell biology, medicine and technology. Angew. Chem. Int. Educ. Eng.,1981,20,325～344
[16]Klenchin. V.A.,S.I. Sukharev, and S.M. Serov. Electrically induced DNA uptake by cells is a fast process involving DNA electrophoresis. Biophys. J.,1991,60,804～811
[17]Sukharev. S.I., V.A. Klencin, and S.M. Serov. Electroporation and electro phoretic DNA transfer into cells, Biophys. J., 1992,63,1320～1372
[18]Wang. M.A.,S.A. Freeman, and V.G. Bose, Theoretical modelling of electro poration: Electrical behavior and molecular transport, In M. Blank, eds. Electricity and Magnetism in Biology and Medicine., San Francisco: San Francisco Press, 1993,138～140
[19]Weaver. J.C., G.I. Harrison and J.G. Bliss. Electroporation: High frequency of occurrence of the transient high permeability state in red blood cells and intact yeast, FEBS Lett., 1988,229,30～34
[20]Sale. A.J.H. and W.A. Hamilton. Effects of high electric fields on microorganisms, I, Killing of bacteria and yeasts. Biochem. biophys. Acta., 1967,148,781～788
[21]Lee. R.C.,and M.S. Kolodney, Electrical injury mechanisms: Electrical breakdown of cell membranes. Plast. Reconstr. Surg., 1987,80,672～679
[22]Lee. R.C.,L.P. River, and F.S. Pan, Surfactant induced sealing of electro permeabilized skeletal muscle membranes in vivo. PNPS.,1992,89,4524～4528
[23]张锦珠，孙彤，陈雅．脉冲电场引起的红血球内钠离子浓度变化的研究．中国科学，C辑，1997，vol．27，no，6：481～485
[24]陈雅，王彦，孙彤．脉冲电场对粒细胞内Ca~(2+)浓度影响的动态变化．中国科学，C辑，1999，vol，29，no．6：663～667
[25]张弘，王保义，刘长军等．利用弱电磁脉冲对细胞电穿孔提高抗癌药物的疗效研究．电波科学学报，1999，14(增刊)：397～400
[26]包家立，梁文权，胡巧红等．经皮给药电穿孔仪的研制。仪器仪表学报，2000，vol．21，no．1：66～69，
[27]J. Mankowski,M. Kristiansen. A review of shot pulsegenerator technology. IEEE Trans. Plasma Sci. 2000, vol. 28, no.1:102～108
[28]Bio-Rad公司产品手册．新一代转基因系统，1999，161～163
[29] 袁其平，徐保强.电脉冲基因导入的实现，见第二届电磁辐射与健康国际研讨会文集 , 2000, 205-206
[30] Dardanoni.L. ,Torregrossa M. V. and Zanforlin. L. Millimeter wave effects on Candida albicans cells, J. Bioelectr. , 1985, 4, 171-176
[31] Levina M.Z.,Veselago.I.A.and Belaya.T.I. Influence of low-intensity VHF irradiation on growth and development of protozoa cultures. In Deyatkav N. D.,eds. Mi11imeter waves in Medicine and Biology. Moscow: Radio electronica, 1989,189-195
[32] Shestopalova. N. G. , Makarenko. B. I. and Golovina. L. N. Modification of synchronizing effects of millimeter wave on first mitoses by different temperature regimens of germination .Millimeter waves in Medicine and Biology. Moscow: IRERAN, 1995, 236-237
[33] Tambiev.A.,Kirikova.N.N. and Lapshin. O. M. The combined effects of exposure to EMF of millimeter waves and centimeter wavelength ranges on productivity of microalgae. In Deyatkav N.D.,eds. Millimeter waves in Medicine and Biology. Moscow: Radio electronica, 1989,183-188
[34] Belyaev. I. Y. ,Alipov.Y.D. and Polunin.V.A. Evidence for dependence of resonant frequency of millimeter wave interaction with Escherichia coli K12 cells on haploid genome length. Electromagnetobiology, 1993, 12:39-49
[35] Rojavin.M.A.,Ziskin.M.C., Effect of millimeter waves on survival of UVC exposed Escherichia coli. Bioelectromagnetics, 1995,16:188-196
[36] Pakhomova.O.N.,Pakhomova.A. G. and Akyel.Y. Effect of mi11imeter waves on UV-induced recombination and mutagenesis in yeast. Bioelectrochem. Bioenerg, 1997,43:227-232
[37] Brovkovich.V.M. Kurilo.N.B. and Barishpol.V. L. Action of millimeter range electromagnetic radiation on the Ca2+ pump of sarcoplasmic reticulum. Radiobiologiia, 1991, 31:268-271
[38] Chernyakov. G. M. , Korochkin. V. L. and Babenko. A. P. reaction of biological system of various complexity to the action of low-level EHF radiation. In Deyatkav N.D., eds. Millimeter waves in Medicine and Biology. Moscow: Radio electronica, 1989,141-167
[39] Burachas.G. ,Mascoliunas.R. Suppression of nerve action potential under the effect of millimeter waves. In Deyatkav N.D.,eds. Millimeter waves in Medicine and Biology. Moscow: Radio electronica, 1989,168-175
[40] GoLant.M.B. Resonance effect of coherent millimeter-band electro magnetic waves on living organisms. Biofizika, 1989, 34:1004-1014
[41]Potekhina. I.L.,Akoyev. G.N. and Oleyner. V.D. Effects fo low-intensity electromagnetic radiation in the millimeter range on the cardio vascular system of the white rat. Fiziol, 1992,78:35～41
[42]Belyaev. I.Y., Okladnova. O.V. and Izmailov. D.M. Differential sensitivity of development stages to low-level electromagnetic radiation of extremely ultrahigh frequency. Dokl Akad Nauk SSSR, 1990,12:68～70
[43]Frei. M.R.,Ryan. K.L. Circulatory failure resulting from sustained millimeter wave irradiation. Brooks AFB, TX:Infrared Lasers and Millimeter Waves Workshop, 1997
[44]Frei. M.R.,Ryan. K.L. Sustained 35GHz radio frequency irradiation induces circulatory failure. Shock, 1995,4:289～293
[45]Temuriants. N.A.,Chuyan. E.N. and Tumaniants. E.N. Dependence of anti-stress effects of millimeter-range electromagnetic fields on localization of exposed area in rats with different typological features. Millimetrovie biologi Medicine, 1993,2:51～59
[46]Frohlich. H. The Biological effects of microwaves and related questions. Adv Electronics Electron Phys, 1980,53:85～152
[47]Frohlich. M. Biological coherence and response to external stimuli. Berlin:Springer-Verlag, 1989,265～268
[48]张富鑫，林崇文．极高频生物医学电子学．成都：电子科技大学出版社，1993
[49]张富鑫，林崇文．极高频治疗学&EHF毫米波多功能治疗仪．成都：电子科技大学出版社，1996
[50]张富鑫．现代信息医学&毫米波临床应用．成都：电子科技大学出版社，2000
[51]吴为民，夏勇，许东卫等．一种采用智能功率模块的调压调频高压电源．高电压技术，2000，vol．26，no．1：66-68
[52]张立，黄两一．电力电子场控器件及其应用．北京：机械工业出版社，1995
[53]黄俊，王兆安．电力电子变流技术．北京：机械工业出版社，1993
[54]张占松，蔡宣三．开关电源的原理与设计．北京：电子工业出版社，2000
[55]汤世贤．微波测量．北京：国防工业出版社，1991
[56]辛华．细胞生物学实验．北京：科学出版社，2001
[57]赵卫红，寿好长，闫福岭．细胞凋亡．河南：河南医科大学出版社，1997
[58]陈念永．毫米波细胞生物效应及抗肿瘤研究：[博士学位论文]．成都：电子科技大学高能所，2001
[2] Abidor. I. G, V.B.Arakelyan, L. V. Chernomordik and Y. A. Chizmadzhev. Electric breakdown of bilayer membranes,I. The main experimental facts and their qualitative discussionBioelectrochem.Bioenerg, 1979,6,37-52
[3] Barnett.A,and J. C. Weaver. Electroporation: A unified quantitative theory of reversible electrical breakdown and rupture. Bioelectrochem. Bioenerg. , 1991, 25, 163-182
[4] Weaver.J.C. Molecular basis for cell membrane electroporation. Proc.N. Y. Acad. Sci., 1993,720,141-152
[5] Chang.D.C, B. M.Chassy and J. A. Saunders. Guide to Electroporation and Electrofusion. Academic,San Diego,Calif,1992
[6] Lipowsky. R. The conformation of membranes. Nature, 1991, 349, 475-481
[7] Pastushenko. V.F. and Y. A. Chizmadzhev. Stabilization of conducting pores in BLM by electric current. 1982,1,43-52
[8] Powell. K. T. and J. C. Weaver. Transient aqueous pores in bilayer membranes: A statistical theory. Bioelectrochem.Bioelectroenerg. , 1986, 15, 211-227
[9] Sabelnikov.A.G. and E.S.Cymbalyuk. Clotting of donor DNA during electroporation of prokaryotes. Bioelectrochem. Bioenerg. 1990,24, 313-321
[10] Crowley.J.M. Electrical breakdown of bimolecular lipid membranes as an electromechanical instability. Biophys,1973,13,711-724
[11] Weaver.J.C. and R.A.Mintzer. Decreased bilayer stability due to transmembrane potentials. Phys. Lett. , 1981,86,57-59
[12] Suarg.I.P. The effects of external fields on the structure of lipid bilayers. J. Physiol. Paris. , 1981,77,1035-1042
[13] Weaver.J.C. and A.Barnett. Progress towards a theoretical model of electroporation mechanism: Membrane electrical behavior and molecular transport. In D. C. Chang, B. M. Chassy, J. A. Saunders, and A. E. Sowers, eds. Guide to electroporation and electrofusion . San Diego: Acdemic, 1992, 91-117
[14]Benz. R. and U. Zimmermann, Relaxation studies on cell membranes and lipid bilayers in high electric field range. Bioelectrochem. bioenerg.,1980,7, 723～739
[15]Zimmermann. U., P. Scheurich, G. Pilwat, and R. Benz Cells with manipulated function: New perspective for cell biology, medicine and technology. Angew. Chem. Int. Educ. Eng.,1981,20,325～344
[16]Klenchin. V.A.,S.I. Sukharev, and S.M. Serov. Electrically induced DNA uptake by cells is a fast process involving DNA electrophoresis. Biophys. J.,1991,60,804～811
[17]Sukharev. S.I., V.A. Klencin, and S.M. Serov. Electroporation and electro phoretic DNA transfer into cells, Biophys. J., 1992,63,1320～1372
[18]Wang. M.A.,S.A. Freeman, and V.G. Bose, Theoretical modelling of electro poration: Electrical behavior and molecular transport, In M. Blank, eds. Electricity and Magnetism in Biology and Medicine., San Francisco: San Francisco Press, 1993,138～140
[19]Weaver. J.C., G.I. Harrison and J.G. Bliss. Electroporation: High frequency of occurrence of the transient high permeability state in red blood cells and intact yeast, FEBS Lett., 1988,229,30～34
[20]Sale. A.J.H. and W.A. Hamilton. Effects of high electric fields on microorganisms, I, Killing of bacteria and yeasts. Biochem. biophys. Acta., 1967,148,781～788
[21]Lee. R.C.,and M.S. Kolodney, Electrical injury mechanisms: Electrical breakdown of cell membranes. Plast. Reconstr. Surg., 1987,80,672～679
[22]Lee. R.C.,L.P. River, and F.S. Pan, Surfactant induced sealing of electro permeabilized skeletal muscle membranes in vivo. PNPS.,1992,89,4524～4528
[23]张锦珠，孙彤，陈雅．脉冲电场引起的红血球内钠离子浓度变化的研究．中国科学，C辑，1997，vol．27，no，6：481～485
[24]陈雅，王彦，孙彤．脉冲电场对粒细胞内Ca~(2+)浓度影响的动态变化．中国科学，C辑，1999，vol，29，no．6：663～667
[25]张弘，王保义，刘长军等．利用弱电磁脉冲对细胞电穿孔提高抗癌药物的疗效研究．电波科学学报，1999，14(增刊)：397～400
[26]包家立，梁文权，胡巧红等．经皮给药电穿孔仪的研制。仪器仪表学报，2000，vol．21，no．1：66～69，
[27]J. Mankowski,M. Kristiansen. A review of shot pulsegenerator technology. IEEE Trans. Plasma Sci. 2000, vol. 28, no.1:102～108
[28]Bio-Rad公司产品手册．新一代转基因系统，1999，161～163
[29] 袁其平，徐保强.电脉冲基因导入的实现，见第二届电磁辐射与健康国际研讨会文集 , 2000, 205-206
[30] Dardanoni.L. ,Torregrossa M. V. and Zanforlin. L. Millimeter wave effects on Candida albicans cells, J. Bioelectr. , 1985, 4, 171-176
[31] Levina M.Z.,Veselago.I.A.and Belaya.T.I. Influence of low-intensity VHF irradiation on growth and development of protozoa cultures. In Deyatkav N. D.,eds. Mi11imeter waves in Medicine and Biology. Moscow: Radio electronica, 1989,189-195
[32] Shestopalova. N. G. , Makarenko. B. I. and Golovina. L. N. Modification of synchronizing effects of millimeter wave on first mitoses by different temperature regimens of germination .Millimeter waves in Medicine and Biology. Moscow: IRERAN, 1995, 236-237
[33] Tambiev.A.,Kirikova.N.N. and Lapshin. O. M. The combined effects of exposure to EMF of millimeter waves and centimeter wavelength ranges on productivity of microalgae. In Deyatkav N.D.,eds. Millimeter waves in Medicine and Biology. Moscow: Radio electronica, 1989,183-188
[34] Belyaev. I. Y. ,Alipov.Y.D. and Polunin.V.A. Evidence for dependence of resonant frequency of millimeter wave interaction with Escherichia coli K12 cells on haploid genome length. Electromagnetobiology, 1993, 12:39-49
[35] Rojavin.M.A.,Ziskin.M.C., Effect of millimeter waves on survival of UVC exposed Escherichia coli. Bioelectromagnetics, 1995,16:188-196
[36] Pakhomova.O.N.,Pakhomova.A. G. and Akyel.Y. Effect of mi11imeter waves on UV-induced recombination and mutagenesis in yeast. Bioelectrochem. Bioenerg, 1997,43:227-232
[37] Brovkovich.V.M. Kurilo.N.B. and Barishpol.V. L. Action of millimeter range electromagnetic radiation on the Ca2+ pump of sarcoplasmic reticulum. Radiobiologiia, 1991, 31:268-271
[38] Chernyakov. G. M. , Korochkin. V. L. and Babenko. A. P. reaction of biological system of various complexity to the action of low-level EHF radiation. In Deyatkav N.D., eds. Millimeter waves in Medicine and Biology. Moscow: Radio electronica, 1989,141-167
[39] Burachas.G. ,Mascoliunas.R. Suppression of nerve action potential under the effect of millimeter waves. In Deyatkav N.D.,eds. Millimeter waves in Medicine and Biology. Moscow: Radio electronica, 1989,168-175
[40] GoLant.M.B. Resonance effect of coherent millimeter-band electro magnetic waves on living organisms. Biofizika, 1989, 34:1004-1014
[41]Potekhina. I.L.,Akoyev. G.N. and Oleyner. V.D. Effects fo low-intensity electromagnetic radiation in the millimeter range on the cardio vascular system of the white rat. Fiziol, 1992,78:35～41
[42]Belyaev. I.Y., Okladnova. O.V. and Izmailov. D.M. Differential sensitivity of development stages to low-level electromagnetic radiation of extremely ultrahigh frequency. Dokl Akad Nauk SSSR, 1990,12:68～70
[43]Frei. M.R.,Ryan. K.L. Circulatory failure resulting from sustained millimeter wave irradiation. Brooks AFB, TX:Infrared Lasers and Millimeter Waves Workshop, 1997
[44]Frei. M.R.,Ryan. K.L. Sustained 35GHz radio frequency irradiation induces circulatory failure. Shock, 1995,4:289～293
[45]Temuriants. N.A.,Chuyan. E.N. and Tumaniants. E.N. Dependence of anti-stress effects of millimeter-range electromagnetic fields on localization of exposed area in rats with different typological features. Millimetrovie biologi Medicine, 1993,2:51～59
[46]Frohlich. H. The Biological effects of microwaves and related questions. Adv Electronics Electron Phys, 1980,53:85～152
[47]Frohlich. M. Biological coherence and response to external stimuli. Berlin:Springer-Verlag, 1989,265～268
[48]张富鑫，林崇文．极高频生物医学电子学．成都：电子科技大学出版社，1993
[49]张富鑫，林崇文．极高频治疗学&EHF毫米波多功能治疗仪．成都：电子科技大学出版社，1996
[50]张富鑫．现代信息医学&毫米波临床应用．成都：电子科技大学出版社，2000
[51]吴为民，夏勇，许东卫等．一种采用智能功率模块的调压调频高压电源．高电压技术，2000，vol．26，no．1：66-68
[52]张立，黄两一．电力电子场控器件及其应用．北京：机械工业出版社，1995
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