人类低品质精液冷冻保存的研究
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摘要
人类低品质精液(Human low-quality semen)，是指精液常规参数异常或／和精子基本功能低下的一大类病理性精液标本。对人类低品质精液进行冷冻保存，是利用一定的低温条件作用于精液，引起精液从生理温度降温、冻结，使精子代谢活动停止而处于休眠状态。人类低品质精液冷冻保存是生殖细胞贮存的一种基本形式，也是人类生殖生物工程当前可以实施的重要技术之一。
本文对低品质精液从病因学来分类，选择4种有代表性的低品质精液类型：(1)精索静脉曲张症(varicocele)患者精液标本；(2)血精症(hematospermia)患者精液标本：(3)圆头精子症(globozoospermia)患者精液标本；(4)精子免疫不育(sperm immunological infertility)患者精液标本，探讨其精子低温生物学特性，冷冻处理方法效果和冷冻精液的临床应用。这4种低品质精液的冷冻保存，国内外未见类似报道。
对精索静脉曲张症精液冷冻保存，取得的主要结果是(1)系统地评价了精索静脉曲张症冷冻精子的动力学、膜完整性、存活时间、顶体蛋白酶活性、穿透宫颈粘液能力的变化；(2)首次阐明精索静脉曲张症冷冻精子受精能力的体外变化特点。这类冷冻精子在解冻后4-6h达到受精能力峰值，但随后迅速下降，保持有效受精能力为8h；(3)探讨了冷冻精子受精能力改变的机理，可能是冷冻解冻过程修饰了精子顶体膜、渗透压效应和易发生获能使受精能力峰值提前；(4)冻贮精索静脉曲张症标本处理后，冷冻精液人工授精取得成功。
对血精症精液冷冻保存，取得的主要结果是：(1)按常规方法冷冻保存精液冷冻效果较差，其原因与血精中凝血效应增大了精液粘稠度不利于精子冷冻存活有关；(2)首创“肝素+精子分离”法处理血精标本，改善了冷冻
Human low-quality semen is a pathological semen, which has low semen routine parameters, characteristics and/or poor sperm functions. During cryopreservation, sperm undergo dramatic changes in intracellular and extracellutar environments due to exposure to cryoprotants, cooling, freezing, storing in liquid nitrogen, and thawing. The chemical and physical effects of these processes may cause extensive cryodamage to sperm membranes that can greatly influence sperm functions for human low-quality semen. Therefore, studying on cryopreservation of human low-quality semen can not only understand changes of frozen-thawed sperm functions, but also promote the application of frozen-thawed low-quality semen.
The present investigation was carried out to evaluate the cryopreservation of four types of low-quality semen, which were from varicocele men, hematospermic men, globozoospermic men, and men with sperm immunological infertility, respectively. These four types semen were selected and divided according to their pathological reasons. For each type of the low-quality semen, the study focused on: (1) analyzing the cryobiological characteristics of frozen thawed sperm; (2) comparing different cryopreservation or treatment techniques; and (3) observing the application of frozen-thawed sperm by using artificial insemination. The obtained results are as follows.
For cryopreservation of semen from men with varicocele, the results showed that: (1) all parameters including motility, membrane integrity rate of HOS-EY test survival time in vitro, cervical mucus penetration ability of SMMPT, acrosomal proteinase activity of frozen-thawed sperm were significantly lowered as compared with those of fresh sperm (P<0.01); (2) the characteristic of
本文对低品质精液从病因学来分类，选择4种有代表性的低品质精液类型：(1)精索静脉曲张症(varicocele)患者精液标本；(2)血精症(hematospermia)患者精液标本：(3)圆头精子症(globozoospermia)患者精液标本；(4)精子免疫不育(sperm immunological infertility)患者精液标本，探讨其精子低温生物学特性，冷冻处理方法效果和冷冻精液的临床应用。这4种低品质精液的冷冻保存，国内外未见类似报道。
对精索静脉曲张症精液冷冻保存，取得的主要结果是(1)系统地评价了精索静脉曲张症冷冻精子的动力学、膜完整性、存活时间、顶体蛋白酶活性、穿透宫颈粘液能力的变化；(2)首次阐明精索静脉曲张症冷冻精子受精能力的体外变化特点。这类冷冻精子在解冻后4-6h达到受精能力峰值，但随后迅速下降，保持有效受精能力为8h；(3)探讨了冷冻精子受精能力改变的机理，可能是冷冻解冻过程修饰了精子顶体膜、渗透压效应和易发生获能使受精能力峰值提前；(4)冻贮精索静脉曲张症标本处理后，冷冻精液人工授精取得成功。
对血精症精液冷冻保存，取得的主要结果是：(1)按常规方法冷冻保存精液冷冻效果较差，其原因与血精中凝血效应增大了精液粘稠度不利于精子冷冻存活有关；(2)首创“肝素+精子分离”法处理血精标本，改善了冷冻
Human low-quality semen is a pathological semen, which has low semen routine parameters, characteristics and/or poor sperm functions. During cryopreservation, sperm undergo dramatic changes in intracellular and extracellutar environments due to exposure to cryoprotants, cooling, freezing, storing in liquid nitrogen, and thawing. The chemical and physical effects of these processes may cause extensive cryodamage to sperm membranes that can greatly influence sperm functions for human low-quality semen. Therefore, studying on cryopreservation of human low-quality semen can not only understand changes of frozen-thawed sperm functions, but also promote the application of frozen-thawed low-quality semen.
The present investigation was carried out to evaluate the cryopreservation of four types of low-quality semen, which were from varicocele men, hematospermic men, globozoospermic men, and men with sperm immunological infertility, respectively. These four types semen were selected and divided according to their pathological reasons. For each type of the low-quality semen, the study focused on: (1) analyzing the cryobiological characteristics of frozen thawed sperm; (2) comparing different cryopreservation or treatment techniques; and (3) observing the application of frozen-thawed sperm by using artificial insemination. The obtained results are as follows.
For cryopreservation of semen from men with varicocele, the results showed that: (1) all parameters including motility, membrane integrity rate of HOS-EY test survival time in vitro, cervical mucus penetration ability of SMMPT, acrosomal proteinase activity of frozen-thawed sperm were significantly lowered as compared with those of fresh sperm (P<0.01); (2) the characteristic of
引文
1. Keel BA, Webster BW. Semen cryopreservation methodology and results. In: Donor Insemination. Barratt CLR, Looke ID, eds. Cambridge University Press, 1993: 71-96.
2．卢光琇．人工授精的发展——冷冻精子库．见：人类生殖与生殖工程．卢惠霖编著，湖南教育出版社，1986：117-130．
3. Hafez ESE. Human Reproduction: Conception and Contraception. Harper and Row Publishers, Inc. 1980: 92-120.
4．郭海禄，胡礼泉主编．临床男科学．湖北科技出版社，1996：15-27．
5．王一飞主编．生殖医学．人民卫生出版社，1991：231-233
6．朱伟杰．人类精子功能检测的现状．暨南大学学报，1995，16(1)：106-110．
7. Polge C, Smith Au, Darkes AS. Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature, 164(15): 666
8. Sherman JK. Research on frozen human semen, Fertil Steril, 1964, 15(5): 485-499
9. Sherman JK. Synopsis of the use of frozen human semen since 1964: State of the art of human semen banking. Fertil Steril, 1973, 24(5): 397-411.
10. Sherman JK. Human semen cryobanks. In: Aspects of Male Infertility. White RV, ed., Willams and Wilkins, 1982: 263-277.
11. Sherman JK. Cryopreservation of human semen. In: CRC Hand book of the Laboratory Diagnosis and Treatment of Infertility. Keel BA, Webster BW, eds., CRC Press, 1990: 229-260.
12. Steinberger S, Smith KD. Artificial insemination with fresh or frozen semen. JAMA, 1973, 223(7): 773-783.
13. Traina G, Traina V. Results of AID with fresh and frozen semen: 488??cases. In: Human Artificial Insemination and Semen Preservation. Devid G, Price WS, eds., Plenum Press, 1980:273-276.
14. Richter MA, Haning RV, Shapiro SS. Artificial donor insemination: fresh versus frozen semen; the patient as her own control. Fertil Steril, 1984, 41(2):277-280.
15. Sato H, Kobayashi T, Mochimaru F, et al. Results of AIH in 1475 patients. In: Human Artificial Insemination and Semen Preservation. David G, Price WS, eds., Plenum Press, 1980: 521-528.
16. Giwercman A, Carlsen E, Keiding N, et al. Evidence for increasing incidence of abnormalities of human testis: A review. Environmental Health Perspectives, 1993, 101(suppl 2): 65-21.
17. Olsen GW, Ross ER, Bodner KM, et al. Have sperm counts been redueed 50 percent in 50 years? A statistical model revisited. Fertil Steril, 1995, 63(4):887-892.
18. Fisch H, Goluboff ET. Geographic variations in sperm counts: a potential cause of bias in studies of semen quality. Fertil Steril, 1996, 65(5):1044-1046.
19. WHO, Enviromental factors blamed for declining sperm quantity and quality. Progress in Hum Reprod Res, 1991, 20:4
20. Palermo G, Joris H, Devroey P, et al. Pregnancies after intracyto- plasmic injection of single spermatozoon into oocyte. Lancet, 1992, 340:17-18.
21. Palermo G, Cohen J, Rosenwaks Z. Intracytoplasmic sperm injection: a powerful tool to overcome fertilization failure. Fertil Steril, 1996, 65(5):899-908.
22. Byrd W, Maddox MA, Ackerman GE, et al. Comparison of bicarbonate and HEPES-buffered media on pregnancy rates after intraterine insemination with cryopreserved donor sperm. Fertil Steril, 1991, ??56(3): 540-546.
23. Sawetawan C, Bruns ES, Prins GS. Improvement of post-thaw sperm motility in poor quality human semen. Fertil Steril, 1993, 60(4): 706-710.
24. Check D J, Check ML, Bollendorf A, et al. Fresh versus frozen seminal plasma for enhancing sperm motility in asthenozoospermic males. Arch Androl, 1990, 26(1): 79-61.
25. Check D J, Check ML, Bollendrof A, et al. Falilure of the addition of fresh seminal plasma to cryoporeservated-thawed sperm to improve semen parameters. Arch Androl, 1993, 31(2): 121-125.
26. Kaneko S, Kobayashi T, Lee HK, et al. Cryogenic preservation of low-quality human semen. Arch Androl, 1990, 24(1): 81-86.
27. Larson JM, Mckinncy KA, Mixon BA, et al. An intrauterine insemina-tion-ready cryopreservation method compared with sperm recovery after conventional freezing and post-thaw proccessing. Fertil Steril,, 1997, 68(1): 143-148.
28. Mbizvo MT, Johnston RC, Backer GHW. The effect of the motility stimulants, caffeine, pentoxifylline, and 2-deoxyad-enosine on hyperactivation of cryopreserved human sperm. Fertil Steril, 1993, 59(5): 1112-1117.
29. Esteves SC, Sharma R, Thomas A, et al. Cryopreservation of human spermatozoa with pentoxifylline improves the post-thaw agonistinduced acrosome reaction rate. Hum Reprod, 1998, 13(12): 3384-389.
30．卢光琇，戴中原，朱敬璋等．人类精液冷冻贮存及临床应用效果的初步报告．湖南医学院学报，1983，8(2)：219-221．
31．朱敬璋，陈涤瑕，徐冬梅等，人类精液冷冻贮存及人工授精的研究．中华医学杂志，1984，64(12)：725-727．
32．刘金刚，王瑞东，葛凤英等．人类精子库及冻精人工授精．青岛医学??院学报，1987，23(2)：8-11．
33. Zhu Wei-Jie(朱伟杰), Liu XG, Li SQ, et al. Human sperm cryobiological characteristics: Comparative cryopreservation techniques. Arch Androl, 1990, 24(1): 95-96.
34．朱伟杰，刘学高，李世勤等．四种人类精液冷冻保护剂的比较研究．中华泌尿外科杂志，1990，11(5)：315-317．
35．朱伟杰，刘学高，李庆琨等．无卵黄型人类精液冷冻保护剂试与临床初步应用．暨南大学学报，1990，11(3)：71-75．
36．于新德，尤国才，徐正铨等．冷冻保存的人类精子超微结构变化．男性学杂志，1995，9(3)：148-151．
37．朱伟杰，刘学高，李世勤等．人类精液的程控冷冻法与悬吊冷冻法比较研究．男性学杂志，1990，4(4)：206-209．
38．何其久，阮圣恒，陈两珍等．冻存精液作人工授精．中华妇产科杂志，1987，22(5)：278-280．
39．梁小薇，沈和，谷立群等．冷冻精液人工授精成功率及其影响因素．生殖医学杂志，1995，4(3)：153-156．
40．王沛涛，崔向东，李冰清．冷冻复温后处理对精子功能影响的研究．男性学杂志，1996，10(1)：1-4．
41. Zhu Wei-Jie(朱伟杰), Liu XG. Cryopreservation of low-quality human semen. In: International Congress on Therapy in Andrology. Fabris GF, Camale D, eds, Monduzzi Editore, 1991: 251-254.
42．朱伟杰，刘学高．人类低品质精液一步冷冻的初步研究．中国病理生理杂志，1992，8(1)：107．
43．郭忠伟，钱宪明，王益鑫等．不育精液冷冻及解冻后上游处理对精子存活率的影响．男性学杂志，1997，11(4)：210-213．
44．李满，庄广伦，李蓉等．冻融附睾精子卵浆内显微注射治疗阻塞性无精子症不育二例．生殖医学杂志，1999，8(1)：53-54。
45．江观泉，于水强主编，雄(男)性生殖毒理学．北京医科大学、中国协??协和医科大学联合出版社，1994：57-63．
46. WHO. Laboratory manual for the examination of human semen and semen-cervical mucus interaction. Cambridge University Press, 1992.
47. Sherman JK. Frozen semen: effeciency in artificial insemination and advantage in testing for acquired immune deficiency syndrome. Fertil Steril, 1987, 47(1): 19-21.
48. Stewart G J, Tyler JPP, Cunningham AL, et al. Transmission of human T-lymphotropic virus typelII(HTLV-Ⅲ) virus by artificial insemination by donor. Lancet, 1985, ⅱ: 581-584.
49. American Fertility Society. Guidelines for human andrology laboratories. Fertil Steril, 1992, 58(Suppl): 1ls-16s.
50．朱伟杰．人类生殖临床精液性STD传播的预防．见：性传播疾病综合防治．中国科协编，中国科学技术出版社，1991：116-119．
51．朱伟杰，刘学高．人类生殖临床精液性STD传播的危险性与我国预防的对策．中国优生优育杂志，1991，2(2)：75-78．
52．国家自然科学基金委员会．1997年度国家自然科学基会项目指南．高等教育出版社，1996；64．
53．国家自然科学基会委员会．1999年度国家自然科学基金申请指南．高等教育出版社，1998：24．
54. 国家重点基础研究发展规划，基础科学研究快报，1999，增刊：8
55. WHO. WHO manual for the standardized investigation and diagnosis of the infertile couple. Rowe PJ, Combaire FH, Hargrave TB, eds., Cam bridge University Press, 1993: 4-39.
56．吴明章，曾超文，张君群主编．男性生殖病理学．上海科学普及出版社，1997：209-220．
57．朱伟杰．人类冷冻精液的巴氏染色．中国临床医药．检验与实验医学．1995，2：1295-1296．
58. Zhu Wei-Jie(朱伟杰), Liu XG. The measurment of membrane integrity??in frozen human sperm by the hypoosmotic swelling-eosin-Y test and its relationship to frozen sperm motility and hamster oocyte penetration assay. Andrologiia, 1996, 4(2): 15-20.
59．朱伟杰，刘学高．冷冻保存对人类精子膜完整性的影响．中国病理生理杂志，1997，13(6)：624-627．
60. Welker B, Bernstein GS, Diedrich K, et al. Acrosomal proteinase activity of human sperm and relation of results to semen quality. Hum Reprod, 1988, 3(suppl): 75-78
61．朱伟杰，姚汝华．冷冻保存对人类精子顶体蛋白酶的影响．暨南大学学报，1998，19(3)：105-108
62．世界卫生组织编．人类精液及精液—宫颈粘液相互作用检测手册．人民卫生出版社，1989：13-22，55-57
63．朱伟杰，刘学高．金黄地鼠卵母细胞4℃保存的研究．中国病理生理杂志，1995，11(2)：238-241．
64．朱伟杰，刘学高．金黄地鼠冷冻保存的初步研究．见：广东生物工程研究．林剑、朱伟杰、罗进贤、姚汝华、黄自然等编．暨南大学出版社，1995：26-29．
65. Ahmadi A, Bongso A, Ng SC. Intracytoplasmic injection of human sperm into the hamster oocyte (hamster ICSI assay) as test for fertilizing capacity of the male-factor sperm. J Assist Reprod Genet, 1996, 13(8): 647-651.
66．朱伟杰，金庆骝，李庆琨．无卵黄型冷冻保护剂配合一步冷冻人类精液的临床应用效果观察．国外医学临床生物化学与检测学分册，1995．15(增刊)：234-235
67．李庆琨，文任乾，李世勤，朱伟杰．两种方法冷冻的精液与新鲜精液人工授精的效果观察．见：广东省生物工程学会论文集，刘学高、朱伟杰编，暨南大学出版社，1992：88-91．
68．朱伟杰，刘学高．人类精液一步冷冻法的研究。中国病理生理杂志，??1993, 9(3): 153-156
69. Netto NR. Varicocele. In: Aspects of Male Infertility. White RV, ed, Wiilams and Wilkins, 1982: 70-88.
70．王益鑫编著，男性不育症诊断与治疗．上海科学技术文献出版社，1998：254-283
71. Gorelick JI, Goldsten M. Loss of fertility in men with varicocele patients. Fertil Steril, 1993, 59(2): 613-616.
72. WHO. The influence of varicocele on parameters of fertility in a large group of men presenting to infertility clinics. Fertil Steril, 1992, 57 (4): 1289-1293.
73. Vermenlen A, Vandeweghe M. Improved foetility after varicocele correction: fact or fiction? Fertil Steril, 1984, 42(2): 249-256.
74. Madgar I, Karasik A, Weissenberg R, et al. Controlled trial of high spermatic vein lization for varicocele in infertily men. Fertil Steril, 1995, 63(1): 120-124.
75．梁蔚波，朱伟杰，苏泽轩等．经腹腔镜精索内静脉高位结扎术．广东医学，1996，17(12)：795-796．
76．余庆骝，朱伟杰，梁蔚波等．精索静脉曲张不育患者术后精液品质变化的远期观察．实用泌尿外科杂志，1995，2(1)：5-7
77. Pozza D, Gregori A, Ossanna P, et al. It is useful to operate on adolescent patients affected by left varicocele? J Androl, 15(suppl): 43-46s.
78. Tinga D, Jager S, Bruijnen CA, et al. Fators related to semen improvement and fertility after varicocele operation. Fertil Steril, 1984, 41(1): 404-410.
79. Lenzi A, Gandini L, Bagolan P, et al. Sperm parameters after early left varicocele treatment. Fertil Steril, 1998, 69(2): 347-349.
80. Cockett AT, Takihara H, Cosentino MJ. The varicocele. Fertil Steril,??1984, 41(1): 5-11.
81. Corson SL, Batzer FR. Insemination data on men with varicoceles. Fertil Steril, 1992, 57(5): 1084-1090.
82. Pinto K J, Krovand L, Jarow JP. Varicocele related testicular atrophy and its predictive effect upon fertility. J Urol, 1994, 152(2): 788-790.
83. Zini A, Bukspan M, Berardinucci D, et al. The influence of clinical and subclinical varicocele on testicular volume. Fertil Steril, 1997, 68(4): 671-674.
84. Naftulin BN, Lewis E, Samuels S, et al. Semen quality in varicocele patients is characterized by tapered sperm cell. Fertil Steril, 1991, 56(1): 149-151.
85．会庆骝，朱伟杰，梁蔚波等。静索静脉曲张不育患者精子形态学分析．临床泌尿外科杂志，1994，15(4)：41-44．
86. Zhu Wei-Jie (朱伟杰), Jim QL, Liang WB, et al. Changes in sperm parameters of infertile men with varicocele after varicocelectomy. In: Symposium of First National Congress of Andrology with International Participation. Bulgarian Andrology Foundation, 1996: 22-23
87. Parikh FR, Kamat SA, Kodwaney GG, et al. Computer-assisted semen analysis parameters in men with varicocele is surgery helpful? Fertil Steril, 1996, 66(3): 440-445.
88. Barthelemy C, Royere D, Hammahah C, et al. Ultrastructural changes in membranes and acrosome of human sperm during cryopreservation. Arch Androl, 1990, 25(1): 29-40.
89. Brotherton J. Cryopreservation of human semen. Arch Androl, 1990, 25(2): 181-195
90. Fernandes PA, Kredentster JV, McCoshen JA, et al. Quantitative analysis of the effect of freezing on donor sperm motion kinetics. Fertil Steril, 1990, 54(2): 322-327.91．刘金刚．人类精子超低温冻贮技术．见：实用简明男性学．吕德滨主编，哈尔滨出版社，1988：275-276．
92. Keel BA, Webster BW, Roberts DK. Effects of cryopreservation on the motility characteristics of human spermatozoa. J Reprod Fertil, 1987, 81(2): 213-220.
93．安民．家畜精液的液态保存．见：家畜繁殖技术进展．中国科学技术情报所重庆分所编．科学技术文献出版社重庆分社，1979：34-35
94. Check ML, Check JH, Long R. Detrimental effects of cryopreservation on the structural and functional integrity of the sperm membrane. Arch Androl, 1991, 27(1): 155-160.
95. Mahanderen MM. Supravital staining of frozen human spermatozoa. Fertil Steril, 1985, 44(6): 842-843.
96. Zhu Wai-Jie (朱伟杰), Liu XG. Application of hypoosmotic swelling test on frozen-thawed human sperm and its relationship to other frozen- thawed sperm function. Chinese J Andrology, 1992, 6(3 suppl): 111.
97. Jeyendran RS, Van der ven HH, Perez-Pelaez M, et al. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other sperm characteristics. J Reprod Fertil, 1984, 70(3): 219-228.
98. Eliasson R, Treichl L. Supravital staining of human spermatozoa. Fertil Steril, 1971, 22(1): 134-135.
99. Jeyendran RS, Van der Ven HH, Zaneveld LTD. The hypoosmotic swelling test: An update. Arch Androl, 1992, 29(2): 105-106.
100. Ahmadi A, Ng SC. The single sperm curling test, a modified hypoosmotic swelling test, as a potential technique for the selection of viable sperm for intracytoplasmic sperm injection. Fertil Steril, 1997, 68(2): 373-375.
101. Liu J, Compton G, Tsai Y L, et al. High fertilization rate obtained??after intracytoplasmic sperm injection with 100％ nomotile spermatozoa selected by using a simple modified hypoosmotic swelling test. 1997, 68(2): 373-375.
102. Critser JK. Cryopreservation of human spermatozoa. Ⅱ postthaw chronology of motility and zona-free hamster ova penetration. Fertil Steril, 47(6): 980-984.
103．朱伟杰，刘学高．精子膜低渗肿胀试验在人类冷冻精子的应用评价．暨南大学学报，1992，13(1)：103-107．
104. Schill WB. Some disturbances of acrosomal development and function in human spermatozoa. Hum Reprod, 1991, 6(7): 969-978.
105. Schill WB, Peterson E T, Heissler E. The sperm acrosome: Functional and clinical aspects. Hum Reprod, 1988, 3(2): 139-145.
106．张适，胡志刚，周作民等．精子功能检测的研究．Ⅰ．底物膜技术对精子顶体酶的测定．男性学杂志，1989，3(4)：193-197．
107．周元聪，朱洪，林南琴等．蛋白酶抑制剂抑制哺乳类精子顶体蛋白酶活性．生殖与避孕，1993，13(2)：114-119．
108．朱伟杰，梁蔚波，金庆骝，人类睾丸精子和附睾精子的顶体蛋白酶活性．中国病理生理杂志，1998，14(5)：465-468．
109. Snyder MG, Zaneveld LJ. Treatment of cervical mucus with lectins: effect on sperm migration. Fertil Steril, 44(5): 633-637.
110. Mortimer D, Pandya IJ, Sawers RS. Relationship between human sperm motility characteristics and sperm penetration into human cervical mucus in vitro. J Reprod Fertil, 1986, 78(1): 93-102.
111. Keel BA, Webster BW. Correlation of human sperm motlity characteristics with an in vitro cervical mucus penetration test. Fertil Steril , 1988, 49(1): 138-143.
112. Katz DF, Morals P, Samuels SJ, et al. Mechanisms of filtration of morphologically abnormal human sperm by cervical mucus. Fertil??Steril, 1990, 54(3): 513-517.
113. Berberoglugil P, Rodesch C, Englert Y, et al. Abnormal Spermmucus penetration test predicts low in vitro fertilization ability of apparently normal semen. Fertil Steril, 1993, 59(6): 1228-1232.
114. Lambert H, Oversteet JW, Morales P, et al. Sperm capacitation in the human female reproductive tract. Fertii Steril, 1985, 43(2): 325-327.
115. Mortimer D, Mortimer ST, Shu MA, et al. A simplified approach to sperm-cercical mucus interaction testing using a hyaluronate migration test. Hum Reprod, 1990, 5(7): 835-841.
116. Eggert-Kruse W, Tilgen W, Leiwhos G, et al. Prognostic value of in vitro sperm penetration into hormonally standardized human cervical mucus. Fertil Steril, 1989, 51(2): 317-323.
117. Alvarez JG, Lasso JL, BlascoL, et al. Centrifugation of human spermatozoa induces sub-lethal damage; Separation of human spermatozoa from seminal plasma by dextran swim-up procedure without centrifuzation extends their motile life. Hum Reprod, 1993, 8(12): 1087-1092.
118. Yanagimachi R. Mammalian fertilization. In: The Physiology of Reproduction. Knobil E, ed., Raven Press, 1988: 135-185.
119. Wassarman PM. Regulation of mammalian fertilization by zona pellucida glycoproteins. J Reprod Fertil, 1990, 42(suppl): 79-87.
120．谢琪旋，朱伟杰，李莉．不孕妇女血清中卵透明节抗体的检测．暨南大学学报，1998，19(增刊)：52-55
121. YanazimachiR, Yanagimachi H, Rogers BJ. The use of zona-free animal ova as a test system for the assessment of fertilizing capacity of human spermatozoa, Bio Reprod, 1976, 15: 471-476.
122. Morshedi M, Ertunc H, Ochniuzer S, et al. Cryopreservation/thawed semen for in vitro fertilization: results from donors and infertile??patients.
123. Zhu Wei-Jie(朱伟杰), Liu XG, Li SQ, et al. Comparative studies of the fertilizing capacity of frozen and fresh human sperm in vitro. Beijing International Symposium on Reproductive Biology, 1990:45.
124. Schill WB, Kohn FM. Cryobanking of spermatozoa: Is there a continuing need for semen cryopreservation. In: Current Advances in Andrology. Waites GMH, Frick J, eds. Monduzzi Editore, 1997:371-382.
125. Liu DY, Baker HWG. Tests of human sperm function and fertilization in vitro. Fertil Steril, 1992,58(3):465-483.
126. Crister JK, Huse-Benda AR, Aaker DV, et al. Cryopreservation of human spermatozoa. I. Effects of holding procedure and seeding on motility, fertilizability, and acrosome reaction. Fertil Steril, 1987, 47(4):456-663.
127. Rogers BJ, Mammalian sperm capacitation and fertilization in vitro: A critique of methodology. Gemete Research, 1978, 1:165-223.
128. Aitken RJ. Capacitation (group disscusion). Int J Andrology, 1986, (suppl 6):156
129. Aitken RJ. The influence of medium composition, osomolarity and fertilization capacity of human spermatozoa. Int J Andrology, 1983, 6(2):180-195.
130. Mazur P. Freezing of living cells: mechanisms and implications. Am J Physiol, 1984, 247:C125-C142.
131. Englert Y, Henlisz A, Delvigne, A, et al. Is fresh or frozen semen to be used in vitro fertilization with donor sperm? Fertil Steril, 1989, 51(4):661-664.
132. Centola GM, Mattox JH, Raubertas RF. Pregnancy rate after doublversus single insemination with frozen donor semen. Fertil steril, 1990, 54(6): 1089-1092.
133. Kruger TF. Sperm morphologic feactures as a prognostic factor in IVF. Fertil steril, 1986, 46(1): 118-125.
134．文任乾，朱伟杰．人类活动精子的分离方法．暨南大学研究生学报，1988，4(2)：98-103．
135．徐崇，刘莉，陈复等．畸精子症的体外疗法——密度梯度离心法分离高活力精子．男性学杂志，1990，4(2)：73-76．
136. Zhu Wei-Jie(朱伟杰), Liu XG, Yao RH. Cryopreservation and separation of sperm from oligozoospermic men. In: 6th Pacific Rim Biotechclinology Conference. HKSTU 1998: PMB.
137．朱伟杰．人类精液的冷冻与解冻方法．暨南大学研究生学报，1990，6(1)：62-66
138. Woolley DM, Richardson DW. Ultrastructwral injury to human spermatozoa after freezing and thawiag. Jo Reprod Fertil, 1978, 53(3): 389-394.
139．朱伟杰．人类精液冷冻保护剂应用研究概况．男性学杂志，1991．5(1)：54-57．
140. Crister JK, Arneson BW, Hase-Benda AR, et al. Cryopreservation of human spermatozoa, Ⅲ. The effect of cryoprotectants on motility. Fertil Steril, 1988, 50(2): 314-320.
141. Tasdemir I, Tasdemir M, Tavukcuoglu S, et al. Effect of abnormal head morphology on the outcome of intracytoplasmic sperm injection in humans. Hum Reprod. 1997, 12(6): 1214-1218.
142．黄平治．血精．见：实用男性学．天津科技出版社编，天津科技出版社，1988：198
143. HafezEFE. Hemospermia. A rchAndrol, 1999, 42(1): 56.
144．薄隽杰．血精症的病因及诊断．中国男性学杂志，1998，12(3)：187．145。吴宏飞，尤国才，陆之康等。血精症的渗断和治疗．男性学杂志，1992，6(3)：168-170．
146. Carrell DT, Kumeck PH, Peterson CM, et al. A randomized, prospective analysis of five sperm preparation techniques before intrauterine insemination of husband sperm. Fertil Steril, 1998, 6(9): 122-126
147．朱伟杰，刘金荣，姚汝华．卵黄缓冲液4℃保存3种低品质精液后的穿卵率变化．中国优生优育杂志，1998，6(6)：14-15．
148．余惠铭主编．病理生理学．人民卫生出版社，1996：136-138．
149. Storey KB, Storey JM. Freeze tolerance in animals. Physiological Review, 1988, 68(1): 27-84.
150. Hill JA. Effects of soluble products of activated lymphocytes and macrophages (lymphokines and monokines) on human sperm motion parameters, Fertil Steril, 1987, 47(3): 460-466.
151．聂振英，白细胞精子症与男性不育．国外医学计划生育分册，1996，15(1)：18-20
152．朱伟杰，姚汝华．卵黄缓冲液4℃保存人精液后的精子存活时间变化，暨南大学学报，1998，19(增刊)：41-43．
153. Soffer Y, Golan A, Herman A, et al. Prediction of in vitro fertilization outcome by sperm penetration assay with TEST-yolk buffer preincubation. Fertil Steril, 1992. 58(3): 556-562
154. Jacobs BR, Caulfield J, Boldt J. Analysis of (TES CETS and Tris) yolk buffer effects on human sperm. Fertil Steril, 1995, 63(5): 1064-1070.
155. Valencia A, Wens MA, Mcrchant H, et al. Capacitation of human sperm by heparin. Arch Androl, 1988, 12(2): 109-113.
156. Saito K, Kinoshita Y, Kanno H, et al. The role of potassiumion and extracellular alkalization in reinitiation of human spermatozoa preserved in electrolycte-free solution at 4℃. Fertil Steril, 1996,??65(6): 1214-1218.
157. Kruger TF, Menkveld k, stander FSH, et al. Sperm morphologic fectures as a prognostic factor in IVF. Fertil Steril, 1986, 46(6): 1118-1123.
158. Sharma KK, Agarwal A. Sperm quality improvement in crypreserved human semen. J Urology, 1996, 156: 1008-1012.
159. Johnson DE, Confino E, Jeyendran KS. Glass wool column filtration versus mini-Percoll Ggradient for processing poor quality semen samples. Fertil Steril, 1996, 66(2): 459-462.
160. Fulgham DL, Alexander N J, Spermatozoa washing and concentration techniques. In: CRC Handbook of the Laboratory Diagnosis and Treatment of Infertility. Keel BA, webster BW, CRC Press, 1990; 193-212.
161．李宏军．精子体外处理技术．男性学杂志，1994，8(3)：188-190．
162．朱伟杰，刘学高．人类冷冻精液的解冻后处理研究．中国病理生理杂志，1994，10(专刊)：154．
163．营英强，江赛男，董协良．人工授精和传染病．生殖与避孕，1990，10(2)：73-75．
164. Eliasson R. Analysis of semen. In: The Testis. Burger H, ole kretser D, eds, Raven Press, 1981: 381-398.
165. Shanis BS, Check JH, Baker AF. Transmission of Sexually transmitted diseases by donor semen. Arch Androl, 1989, 23(3): 249-257.
166. Greenblatt RM, Handsfield HH, Sayers MH. Screening therapeutic insemination donors for sexually transmitted diseases: overview and recommendation. Fertil Steril, 1986, 43(3): 351-364.
167. Prins GS, Weidel L. A Comparative study of buffer systems as cryoprotectants for human spermatozoa. Fentil Steril, 1986, 46(1): 146-148.168. Weidel L, Prins GS. Cryosurvival of human spermatozoa frozen in eight different buffer systems. J Androl, 1987, 8(1):41-47.
169. Kovacs G, Baker G, Burger H, et al. Artificial insemination with cryopreserved donor semen: a decade of experience. British J Obstet Gynaecol, 1988, 95(3):354-360.
170. Wong AWY, Kan M, Ma HK. Factors affecting the success of artificial insemination by frozen donor semen. Int. J Fertil, 1989, 34(1 ):25-29.
171. Aiman J. Factors affecting the success of donor insemination. Fertil Steril, 1982, 37(1): 94-99.
172. Lamprecht IA, Kotzur B, Schopf E. Round-headed human sperm. Fentil Steril, 1976, 27(6): 685-693.
173. Pederson J, Rebbe H. Fine structure of round-headed human spermatozoa. J Reprod Fertil, 1974, 37(1):51-54.
174. Flork-Gerloff SF, Toper-Peterson E, Muller-Ester. W, et al. Biochemical and genetic investigation of round-headed spermatozoa in infertile men induding two brother and their father. Androlozia, 1984, 16(2):187-202.
175. Check JH, Kozak J, Katsoff D. Hypoosmotic swelling: evalnation of sperm tails using stict criteria in cases of globozoospermia. Arch Androl, 1993, 30(2): 125-128.
176. Lalonde L, Langlais J, Antakl P, et al. Male infertility associated with round-headed acrosomeless spermatozoa. Fertil Steril, 1988, 49(2):316-321.
177. Weissenberg R, Eshkol A, Rudak E, et al. Inability of round-headed acrosomeless human spermatozoa to penetrate zona-free hamster ova. Arch Androl, 1982, 11 (2): 167-169.
178. Syms AJ, Johnson AR, Lip Shultz, LI, et al. Studies on human spermatozoa with round head syndrome. Andrologia, 1984, 42(4):431-435.
179. Rybouchkin AV, Sutter PD, Straeten FV, et al. Fertilization and pregnancy after assisted oocyte activation and intracytoplasmic sperm injection in a case of round-headed sperm associated with deficient oocyte activation capacity. Fertil Steril, 1997, 68(6): 1144-1147.
180. Battaglia DE, Koebler JK, Klein NA, et al. Failure ofoocyte activation after intracytoplasmic sperm injection using round-headed sperm. Fertil Steril, 1997, 68(1): 118-122.
181. Bourne H, Liu DY, Clarke GN, et al. Normal fertilization and embryo development by intracytoplasmic sperm injection of round-headed acrosomeless sperm. Fertil Steril, 1995, 63(6): 1329-1332.
182. Cha K Y, Oum KB, Kim HJ. Approaches for obtaining sperm in patients with male factor infertility, Fertil Steril, 1997, 67(6): 985-995.
183．朱伟杰，刘学高．精子免疫与免疫不育。见：遗传优生与生殖工程．梁志成，朱嘉铭，朱伟杰等编．暨南大学出版社，1992：211-216．
184. Bronson R, Cooper G, Rosenfeld. Sperm antibdies: their role in infertility. Fertil Steril, 1984, 42(2): 171-183.
185. Geva E, Amit A, Lerner-Geval L, et al. Autoimmunity and reproduction. Fertil Steril, 1997, 67(4): 599-611.
186. Kamada M, Yamano S, Senuma M, et al. Semen analysis and antisperm antibody. Arch Androi, 1998, 40(2): 117-128.
187. Naz RK, Menge A. Antisperm antibodies: origin, regulation, and sperm reactivity in human infertility. Fertil Steril, 1994, 61(6): 1001-1013.
188．朱伟杰，梁蔚波，会庆骝等．不育患者睾丸活检前、后的血清抗精子抗体检测．见：广东省生物工程学会论文集．刘学高，朱伟杰编，暨南大学出版社，1992：45-47．
189. Alexander NJ. Treatment for antisperm antibodies: voodoo or victory???Fetil Steril. 1990. 53(4): 602-603.
190．刘继红，章咏裳．免疫与男性不育症．见：生殖免疫学．罗丽兰主编，湖北科技出版社，1998：184-198．
191. Hass GG, Cunningham M, Culp L. The effect of freezing on spermassociated immunoglobulin G(IgG). Fertii Steril, 1984, 42(5): 761-764.
192. Vazquez-Levin MH, Notrica JA, Fried EP. Male immunologic infertility: sperm performance on in vitro fertilization. Fertil Stril, 1997, 68(4): 675-681.
193. Clarke GN, Bourne H, Baker HWG. Intracytoplasmic sperm injection for treating infertility associated with sperm antoimmunity. Fertil Steril, 1997, 68(1): 112-117.
194. Marshburn PB, Kuttch, WH. The role of antisperm antibodies in infertility. Fertil Steril, 1994, 61(5): 799-811.
2．卢光琇．人工授精的发展——冷冻精子库．见：人类生殖与生殖工程．卢惠霖编著，湖南教育出版社，1986：117-130．
3. Hafez ESE. Human Reproduction: Conception and Contraception. Harper and Row Publishers, Inc. 1980: 92-120.
4．郭海禄，胡礼泉主编．临床男科学．湖北科技出版社，1996：15-27．
5．王一飞主编．生殖医学．人民卫生出版社，1991：231-233
6．朱伟杰．人类精子功能检测的现状．暨南大学学报，1995，16(1)：106-110．
7. Polge C, Smith Au, Darkes AS. Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature, 164(15): 666
8. Sherman JK. Research on frozen human semen, Fertil Steril, 1964, 15(5): 485-499
9. Sherman JK. Synopsis of the use of frozen human semen since 1964: State of the art of human semen banking. Fertil Steril, 1973, 24(5): 397-411.
10. Sherman JK. Human semen cryobanks. In: Aspects of Male Infertility. White RV, ed., Willams and Wilkins, 1982: 263-277.
11. Sherman JK. Cryopreservation of human semen. In: CRC Hand book of the Laboratory Diagnosis and Treatment of Infertility. Keel BA, Webster BW, eds., CRC Press, 1990: 229-260.
12. Steinberger S, Smith KD. Artificial insemination with fresh or frozen semen. JAMA, 1973, 223(7): 773-783.
13. Traina G, Traina V. Results of AID with fresh and frozen semen: 488??cases. In: Human Artificial Insemination and Semen Preservation. Devid G, Price WS, eds., Plenum Press, 1980:273-276.
14. Richter MA, Haning RV, Shapiro SS. Artificial donor insemination: fresh versus frozen semen; the patient as her own control. Fertil Steril, 1984, 41(2):277-280.
15. Sato H, Kobayashi T, Mochimaru F, et al. Results of AIH in 1475 patients. In: Human Artificial Insemination and Semen Preservation. David G, Price WS, eds., Plenum Press, 1980: 521-528.
16. Giwercman A, Carlsen E, Keiding N, et al. Evidence for increasing incidence of abnormalities of human testis: A review. Environmental Health Perspectives, 1993, 101(suppl 2): 65-21.
17. Olsen GW, Ross ER, Bodner KM, et al. Have sperm counts been redueed 50 percent in 50 years? A statistical model revisited. Fertil Steril, 1995, 63(4):887-892.
18. Fisch H, Goluboff ET. Geographic variations in sperm counts: a potential cause of bias in studies of semen quality. Fertil Steril, 1996, 65(5):1044-1046.
19. WHO, Enviromental factors blamed for declining sperm quantity and quality. Progress in Hum Reprod Res, 1991, 20:4
20. Palermo G, Joris H, Devroey P, et al. Pregnancies after intracyto- plasmic injection of single spermatozoon into oocyte. Lancet, 1992, 340:17-18.
21. Palermo G, Cohen J, Rosenwaks Z. Intracytoplasmic sperm injection: a powerful tool to overcome fertilization failure. Fertil Steril, 1996, 65(5):899-908.
22. Byrd W, Maddox MA, Ackerman GE, et al. Comparison of bicarbonate and HEPES-buffered media on pregnancy rates after intraterine insemination with cryopreserved donor sperm. Fertil Steril, 1991, ??56(3): 540-546.
23. Sawetawan C, Bruns ES, Prins GS. Improvement of post-thaw sperm motility in poor quality human semen. Fertil Steril, 1993, 60(4): 706-710.
24. Check D J, Check ML, Bollendorf A, et al. Fresh versus frozen seminal plasma for enhancing sperm motility in asthenozoospermic males. Arch Androl, 1990, 26(1): 79-61.
25. Check D J, Check ML, Bollendrof A, et al. Falilure of the addition of fresh seminal plasma to cryoporeservated-thawed sperm to improve semen parameters. Arch Androl, 1993, 31(2): 121-125.
26. Kaneko S, Kobayashi T, Lee HK, et al. Cryogenic preservation of low-quality human semen. Arch Androl, 1990, 24(1): 81-86.
27. Larson JM, Mckinncy KA, Mixon BA, et al. An intrauterine insemina-tion-ready cryopreservation method compared with sperm recovery after conventional freezing and post-thaw proccessing. Fertil Steril,, 1997, 68(1): 143-148.
28. Mbizvo MT, Johnston RC, Backer GHW. The effect of the motility stimulants, caffeine, pentoxifylline, and 2-deoxyad-enosine on hyperactivation of cryopreserved human sperm. Fertil Steril, 1993, 59(5): 1112-1117.
29. Esteves SC, Sharma R, Thomas A, et al. Cryopreservation of human spermatozoa with pentoxifylline improves the post-thaw agonistinduced acrosome reaction rate. Hum Reprod, 1998, 13(12): 3384-389.
30．卢光琇，戴中原，朱敬璋等．人类精液冷冻贮存及临床应用效果的初步报告．湖南医学院学报，1983，8(2)：219-221．
31．朱敬璋，陈涤瑕，徐冬梅等，人类精液冷冻贮存及人工授精的研究．中华医学杂志，1984，64(12)：725-727．
32．刘金刚，王瑞东，葛凤英等．人类精子库及冻精人工授精．青岛医学??院学报，1987，23(2)：8-11．
33. Zhu Wei-Jie(朱伟杰), Liu XG, Li SQ, et al. Human sperm cryobiological characteristics: Comparative cryopreservation techniques. Arch Androl, 1990, 24(1): 95-96.
34．朱伟杰，刘学高，李世勤等．四种人类精液冷冻保护剂的比较研究．中华泌尿外科杂志，1990，11(5)：315-317．
35．朱伟杰，刘学高，李庆琨等．无卵黄型人类精液冷冻保护剂试与临床初步应用．暨南大学学报，1990，11(3)：71-75．
36．于新德，尤国才，徐正铨等．冷冻保存的人类精子超微结构变化．男性学杂志，1995，9(3)：148-151．
37．朱伟杰，刘学高，李世勤等．人类精液的程控冷冻法与悬吊冷冻法比较研究．男性学杂志，1990，4(4)：206-209．
38．何其久，阮圣恒，陈两珍等．冻存精液作人工授精．中华妇产科杂志，1987，22(5)：278-280．
39．梁小薇，沈和，谷立群等．冷冻精液人工授精成功率及其影响因素．生殖医学杂志，1995，4(3)：153-156．
40．王沛涛，崔向东，李冰清．冷冻复温后处理对精子功能影响的研究．男性学杂志，1996，10(1)：1-4．
41. Zhu Wei-Jie(朱伟杰), Liu XG. Cryopreservation of low-quality human semen. In: International Congress on Therapy in Andrology. Fabris GF, Camale D, eds, Monduzzi Editore, 1991: 251-254.
42．朱伟杰，刘学高．人类低品质精液一步冷冻的初步研究．中国病理生理杂志，1992，8(1)：107．
43．郭忠伟，钱宪明，王益鑫等．不育精液冷冻及解冻后上游处理对精子存活率的影响．男性学杂志，1997，11(4)：210-213．
44．李满，庄广伦，李蓉等．冻融附睾精子卵浆内显微注射治疗阻塞性无精子症不育二例．生殖医学杂志，1999，8(1)：53-54。
45．江观泉，于水强主编，雄(男)性生殖毒理学．北京医科大学、中国协??协和医科大学联合出版社，1994：57-63．
46. WHO. Laboratory manual for the examination of human semen and semen-cervical mucus interaction. Cambridge University Press, 1992.
47. Sherman JK. Frozen semen: effeciency in artificial insemination and advantage in testing for acquired immune deficiency syndrome. Fertil Steril, 1987, 47(1): 19-21.
48. Stewart G J, Tyler JPP, Cunningham AL, et al. Transmission of human T-lymphotropic virus typelII(HTLV-Ⅲ) virus by artificial insemination by donor. Lancet, 1985, ⅱ: 581-584.
49. American Fertility Society. Guidelines for human andrology laboratories. Fertil Steril, 1992, 58(Suppl): 1ls-16s.
50．朱伟杰．人类生殖临床精液性STD传播的预防．见：性传播疾病综合防治．中国科协编，中国科学技术出版社，1991：116-119．
51．朱伟杰，刘学高．人类生殖临床精液性STD传播的危险性与我国预防的对策．中国优生优育杂志，1991，2(2)：75-78．
52．国家自然科学基金委员会．1997年度国家自然科学基会项目指南．高等教育出版社，1996；64．
53．国家自然科学基会委员会．1999年度国家自然科学基金申请指南．高等教育出版社，1998：24．
54. 国家重点基础研究发展规划，基础科学研究快报，1999，增刊：8
55. WHO. WHO manual for the standardized investigation and diagnosis of the infertile couple. Rowe PJ, Combaire FH, Hargrave TB, eds., Cam bridge University Press, 1993: 4-39.
56．吴明章，曾超文，张君群主编．男性生殖病理学．上海科学普及出版社，1997：209-220．
57．朱伟杰．人类冷冻精液的巴氏染色．中国临床医药．检验与实验医学．1995，2：1295-1296．
58. Zhu Wei-Jie(朱伟杰), Liu XG. The measurment of membrane integrity??in frozen human sperm by the hypoosmotic swelling-eosin-Y test and its relationship to frozen sperm motility and hamster oocyte penetration assay. Andrologiia, 1996, 4(2): 15-20.
59．朱伟杰，刘学高．冷冻保存对人类精子膜完整性的影响．中国病理生理杂志，1997，13(6)：624-627．
60. Welker B, Bernstein GS, Diedrich K, et al. Acrosomal proteinase activity of human sperm and relation of results to semen quality. Hum Reprod, 1988, 3(suppl): 75-78
61．朱伟杰，姚汝华．冷冻保存对人类精子顶体蛋白酶的影响．暨南大学学报，1998，19(3)：105-108
62．世界卫生组织编．人类精液及精液—宫颈粘液相互作用检测手册．人民卫生出版社，1989：13-22，55-57
63．朱伟杰，刘学高．金黄地鼠卵母细胞4℃保存的研究．中国病理生理杂志，1995，11(2)：238-241．
64．朱伟杰，刘学高．金黄地鼠冷冻保存的初步研究．见：广东生物工程研究．林剑、朱伟杰、罗进贤、姚汝华、黄自然等编．暨南大学出版社，1995：26-29．
65. Ahmadi A, Bongso A, Ng SC. Intracytoplasmic injection of human sperm into the hamster oocyte (hamster ICSI assay) as test for fertilizing capacity of the male-factor sperm. J Assist Reprod Genet, 1996, 13(8): 647-651.
66．朱伟杰，金庆骝，李庆琨．无卵黄型冷冻保护剂配合一步冷冻人类精液的临床应用效果观察．国外医学临床生物化学与检测学分册，1995．15(增刊)：234-235
67．李庆琨，文任乾，李世勤，朱伟杰．两种方法冷冻的精液与新鲜精液人工授精的效果观察．见：广东省生物工程学会论文集，刘学高、朱伟杰编，暨南大学出版社，1992：88-91．
68．朱伟杰，刘学高．人类精液一步冷冻法的研究。中国病理生理杂志，??1993, 9(3): 153-156
69. Netto NR. Varicocele. In: Aspects of Male Infertility. White RV, ed, Wiilams and Wilkins, 1982: 70-88.
70．王益鑫编著，男性不育症诊断与治疗．上海科学技术文献出版社，1998：254-283
71. Gorelick JI, Goldsten M. Loss of fertility in men with varicocele patients. Fertil Steril, 1993, 59(2): 613-616.
72. WHO. The influence of varicocele on parameters of fertility in a large group of men presenting to infertility clinics. Fertil Steril, 1992, 57 (4): 1289-1293.
73. Vermenlen A, Vandeweghe M. Improved foetility after varicocele correction: fact or fiction? Fertil Steril, 1984, 42(2): 249-256.
74. Madgar I, Karasik A, Weissenberg R, et al. Controlled trial of high spermatic vein lization for varicocele in infertily men. Fertil Steril, 1995, 63(1): 120-124.
75．梁蔚波，朱伟杰，苏泽轩等．经腹腔镜精索内静脉高位结扎术．广东医学，1996，17(12)：795-796．
76．余庆骝，朱伟杰，梁蔚波等．精索静脉曲张不育患者术后精液品质变化的远期观察．实用泌尿外科杂志，1995，2(1)：5-7
77. Pozza D, Gregori A, Ossanna P, et al. It is useful to operate on adolescent patients affected by left varicocele? J Androl, 15(suppl): 43-46s.
78. Tinga D, Jager S, Bruijnen CA, et al. Fators related to semen improvement and fertility after varicocele operation. Fertil Steril, 1984, 41(1): 404-410.
79. Lenzi A, Gandini L, Bagolan P, et al. Sperm parameters after early left varicocele treatment. Fertil Steril, 1998, 69(2): 347-349.
80. Cockett AT, Takihara H, Cosentino MJ. The varicocele. Fertil Steril,??1984, 41(1): 5-11.
81. Corson SL, Batzer FR. Insemination data on men with varicoceles. Fertil Steril, 1992, 57(5): 1084-1090.
82. Pinto K J, Krovand L, Jarow JP. Varicocele related testicular atrophy and its predictive effect upon fertility. J Urol, 1994, 152(2): 788-790.
83. Zini A, Bukspan M, Berardinucci D, et al. The influence of clinical and subclinical varicocele on testicular volume. Fertil Steril, 1997, 68(4): 671-674.
84. Naftulin BN, Lewis E, Samuels S, et al. Semen quality in varicocele patients is characterized by tapered sperm cell. Fertil Steril, 1991, 56(1): 149-151.
85．会庆骝，朱伟杰，梁蔚波等。静索静脉曲张不育患者精子形态学分析．临床泌尿外科杂志，1994，15(4)：41-44．
86. Zhu Wei-Jie (朱伟杰), Jim QL, Liang WB, et al. Changes in sperm parameters of infertile men with varicocele after varicocelectomy. In: Symposium of First National Congress of Andrology with International Participation. Bulgarian Andrology Foundation, 1996: 22-23
87. Parikh FR, Kamat SA, Kodwaney GG, et al. Computer-assisted semen analysis parameters in men with varicocele is surgery helpful? Fertil Steril, 1996, 66(3): 440-445.
88. Barthelemy C, Royere D, Hammahah C, et al. Ultrastructural changes in membranes and acrosome of human sperm during cryopreservation. Arch Androl, 1990, 25(1): 29-40.
89. Brotherton J. Cryopreservation of human semen. Arch Androl, 1990, 25(2): 181-195
90. Fernandes PA, Kredentster JV, McCoshen JA, et al. Quantitative analysis of the effect of freezing on donor sperm motion kinetics. Fertil Steril, 1990, 54(2): 322-327.91．刘金刚．人类精子超低温冻贮技术．见：实用简明男性学．吕德滨主编，哈尔滨出版社，1988：275-276．
92. Keel BA, Webster BW, Roberts DK. Effects of cryopreservation on the motility characteristics of human spermatozoa. J Reprod Fertil, 1987, 81(2): 213-220.
93．安民．家畜精液的液态保存．见：家畜繁殖技术进展．中国科学技术情报所重庆分所编．科学技术文献出版社重庆分社，1979：34-35
94. Check ML, Check JH, Long R. Detrimental effects of cryopreservation on the structural and functional integrity of the sperm membrane. Arch Androl, 1991, 27(1): 155-160.
95. Mahanderen MM. Supravital staining of frozen human spermatozoa. Fertil Steril, 1985, 44(6): 842-843.
96. Zhu Wai-Jie (朱伟杰), Liu XG. Application of hypoosmotic swelling test on frozen-thawed human sperm and its relationship to other frozen- thawed sperm function. Chinese J Andrology, 1992, 6(3 suppl): 111.
97. Jeyendran RS, Van der ven HH, Perez-Pelaez M, et al. Development of an assay to assess the functional integrity of the human sperm membrane and its relationship to other sperm characteristics. J Reprod Fertil, 1984, 70(3): 219-228.
98. Eliasson R, Treichl L. Supravital staining of human spermatozoa. Fertil Steril, 1971, 22(1): 134-135.
99. Jeyendran RS, Van der Ven HH, Zaneveld LTD. The hypoosmotic swelling test: An update. Arch Androl, 1992, 29(2): 105-106.
100. Ahmadi A, Ng SC. The single sperm curling test, a modified hypoosmotic swelling test, as a potential technique for the selection of viable sperm for intracytoplasmic sperm injection. Fertil Steril, 1997, 68(2): 373-375.
101. Liu J, Compton G, Tsai Y L, et al. High fertilization rate obtained??after intracytoplasmic sperm injection with 100％ nomotile spermatozoa selected by using a simple modified hypoosmotic swelling test. 1997, 68(2): 373-375.
102. Critser JK. Cryopreservation of human spermatozoa. Ⅱ postthaw chronology of motility and zona-free hamster ova penetration. Fertil Steril, 47(6): 980-984.
103．朱伟杰，刘学高．精子膜低渗肿胀试验在人类冷冻精子的应用评价．暨南大学学报，1992，13(1)：103-107．
104. Schill WB. Some disturbances of acrosomal development and function in human spermatozoa. Hum Reprod, 1991, 6(7): 969-978.
105. Schill WB, Peterson E T, Heissler E. The sperm acrosome: Functional and clinical aspects. Hum Reprod, 1988, 3(2): 139-145.
106．张适，胡志刚，周作民等．精子功能检测的研究．Ⅰ．底物膜技术对精子顶体酶的测定．男性学杂志，1989，3(4)：193-197．
107．周元聪，朱洪，林南琴等．蛋白酶抑制剂抑制哺乳类精子顶体蛋白酶活性．生殖与避孕，1993，13(2)：114-119．
108．朱伟杰，梁蔚波，金庆骝，人类睾丸精子和附睾精子的顶体蛋白酶活性．中国病理生理杂志，1998，14(5)：465-468．
109. Snyder MG, Zaneveld LJ. Treatment of cervical mucus with lectins: effect on sperm migration. Fertil Steril, 44(5): 633-637.
110. Mortimer D, Pandya IJ, Sawers RS. Relationship between human sperm motility characteristics and sperm penetration into human cervical mucus in vitro. J Reprod Fertil, 1986, 78(1): 93-102.
111. Keel BA, Webster BW. Correlation of human sperm motlity characteristics with an in vitro cervical mucus penetration test. Fertil Steril , 1988, 49(1): 138-143.
112. Katz DF, Morals P, Samuels SJ, et al. Mechanisms of filtration of morphologically abnormal human sperm by cervical mucus. Fertil??Steril, 1990, 54(3): 513-517.
113. Berberoglugil P, Rodesch C, Englert Y, et al. Abnormal Spermmucus penetration test predicts low in vitro fertilization ability of apparently normal semen. Fertil Steril, 1993, 59(6): 1228-1232.
114. Lambert H, Oversteet JW, Morales P, et al. Sperm capacitation in the human female reproductive tract. Fertii Steril, 1985, 43(2): 325-327.
115. Mortimer D, Mortimer ST, Shu MA, et al. A simplified approach to sperm-cercical mucus interaction testing using a hyaluronate migration test. Hum Reprod, 1990, 5(7): 835-841.
116. Eggert-Kruse W, Tilgen W, Leiwhos G, et al. Prognostic value of in vitro sperm penetration into hormonally standardized human cervical mucus. Fertil Steril, 1989, 51(2): 317-323.
117. Alvarez JG, Lasso JL, BlascoL, et al. Centrifugation of human spermatozoa induces sub-lethal damage; Separation of human spermatozoa from seminal plasma by dextran swim-up procedure without centrifuzation extends their motile life. Hum Reprod, 1993, 8(12): 1087-1092.
118. Yanagimachi R. Mammalian fertilization. In: The Physiology of Reproduction. Knobil E, ed., Raven Press, 1988: 135-185.
119. Wassarman PM. Regulation of mammalian fertilization by zona pellucida glycoproteins. J Reprod Fertil, 1990, 42(suppl): 79-87.
120．谢琪旋，朱伟杰，李莉．不孕妇女血清中卵透明节抗体的检测．暨南大学学报，1998，19(增刊)：52-55
121. YanazimachiR, Yanagimachi H, Rogers BJ. The use of zona-free animal ova as a test system for the assessment of fertilizing capacity of human spermatozoa, Bio Reprod, 1976, 15: 471-476.
122. Morshedi M, Ertunc H, Ochniuzer S, et al. Cryopreservation/thawed semen for in vitro fertilization: results from donors and infertile??patients.
123. Zhu Wei-Jie(朱伟杰), Liu XG, Li SQ, et al. Comparative studies of the fertilizing capacity of frozen and fresh human sperm in vitro. Beijing International Symposium on Reproductive Biology, 1990:45.
124. Schill WB, Kohn FM. Cryobanking of spermatozoa: Is there a continuing need for semen cryopreservation. In: Current Advances in Andrology. Waites GMH, Frick J, eds. Monduzzi Editore, 1997:371-382.
125. Liu DY, Baker HWG. Tests of human sperm function and fertilization in vitro. Fertil Steril, 1992,58(3):465-483.
126. Crister JK, Huse-Benda AR, Aaker DV, et al. Cryopreservation of human spermatozoa. I. Effects of holding procedure and seeding on motility, fertilizability, and acrosome reaction. Fertil Steril, 1987, 47(4):456-663.
127. Rogers BJ, Mammalian sperm capacitation and fertilization in vitro: A critique of methodology. Gemete Research, 1978, 1:165-223.
128. Aitken RJ. Capacitation (group disscusion). Int J Andrology, 1986, (suppl 6):156
129. Aitken RJ. The influence of medium composition, osomolarity and fertilization capacity of human spermatozoa. Int J Andrology, 1983, 6(2):180-195.
130. Mazur P. Freezing of living cells: mechanisms and implications. Am J Physiol, 1984, 247:C125-C142.
131. Englert Y, Henlisz A, Delvigne, A, et al. Is fresh or frozen semen to be used in vitro fertilization with donor sperm? Fertil Steril, 1989, 51(4):661-664.
132. Centola GM, Mattox JH, Raubertas RF. Pregnancy rate after doublversus single insemination with frozen donor semen. Fertil steril, 1990, 54(6): 1089-1092.
133. Kruger TF. Sperm morphologic feactures as a prognostic factor in IVF. Fertil steril, 1986, 46(1): 118-125.
134．文任乾，朱伟杰．人类活动精子的分离方法．暨南大学研究生学报，1988，4(2)：98-103．
135．徐崇，刘莉，陈复等．畸精子症的体外疗法——密度梯度离心法分离高活力精子．男性学杂志，1990，4(2)：73-76．
136. Zhu Wei-Jie(朱伟杰), Liu XG, Yao RH. Cryopreservation and separation of sperm from oligozoospermic men. In: 6th Pacific Rim Biotechclinology Conference. HKSTU 1998: PMB.
137．朱伟杰．人类精液的冷冻与解冻方法．暨南大学研究生学报，1990，6(1)：62-66
138. Woolley DM, Richardson DW. Ultrastructwral injury to human spermatozoa after freezing and thawiag. Jo Reprod Fertil, 1978, 53(3): 389-394.
139．朱伟杰．人类精液冷冻保护剂应用研究概况．男性学杂志，1991．5(1)：54-57．
140. Crister JK, Arneson BW, Hase-Benda AR, et al. Cryopreservation of human spermatozoa, Ⅲ. The effect of cryoprotectants on motility. Fertil Steril, 1988, 50(2): 314-320.
141. Tasdemir I, Tasdemir M, Tavukcuoglu S, et al. Effect of abnormal head morphology on the outcome of intracytoplasmic sperm injection in humans. Hum Reprod. 1997, 12(6): 1214-1218.
142．黄平治．血精．见：实用男性学．天津科技出版社编，天津科技出版社，1988：198
143. HafezEFE. Hemospermia. A rchAndrol, 1999, 42(1): 56.
144．薄隽杰．血精症的病因及诊断．中国男性学杂志，1998，12(3)：187．145。吴宏飞，尤国才，陆之康等。血精症的渗断和治疗．男性学杂志，1992，6(3)：168-170．
146. Carrell DT, Kumeck PH, Peterson CM, et al. A randomized, prospective analysis of five sperm preparation techniques before intrauterine insemination of husband sperm. Fertil Steril, 1998, 6(9): 122-126
147．朱伟杰，刘金荣，姚汝华．卵黄缓冲液4℃保存3种低品质精液后的穿卵率变化．中国优生优育杂志，1998，6(6)：14-15．
148．余惠铭主编．病理生理学．人民卫生出版社，1996：136-138．
149. Storey KB, Storey JM. Freeze tolerance in animals. Physiological Review, 1988, 68(1): 27-84.
150. Hill JA. Effects of soluble products of activated lymphocytes and macrophages (lymphokines and monokines) on human sperm motion parameters, Fertil Steril, 1987, 47(3): 460-466.
151．聂振英，白细胞精子症与男性不育．国外医学计划生育分册，1996，15(1)：18-20
152．朱伟杰，姚汝华．卵黄缓冲液4℃保存人精液后的精子存活时间变化，暨南大学学报，1998，19(增刊)：41-43．
153. Soffer Y, Golan A, Herman A, et al. Prediction of in vitro fertilization outcome by sperm penetration assay with TEST-yolk buffer preincubation. Fertil Steril, 1992. 58(3): 556-562
154. Jacobs BR, Caulfield J, Boldt J. Analysis of (TES CETS and Tris) yolk buffer effects on human sperm. Fertil Steril, 1995, 63(5): 1064-1070.
155. Valencia A, Wens MA, Mcrchant H, et al. Capacitation of human sperm by heparin. Arch Androl, 1988, 12(2): 109-113.
156. Saito K, Kinoshita Y, Kanno H, et al. The role of potassiumion and extracellular alkalization in reinitiation of human spermatozoa preserved in electrolycte-free solution at 4℃. Fertil Steril, 1996,??65(6): 1214-1218.
157. Kruger TF, Menkveld k, stander FSH, et al. Sperm morphologic fectures as a prognostic factor in IVF. Fertil Steril, 1986, 46(6): 1118-1123.
158. Sharma KK, Agarwal A. Sperm quality improvement in crypreserved human semen. J Urology, 1996, 156: 1008-1012.
159. Johnson DE, Confino E, Jeyendran KS. Glass wool column filtration versus mini-Percoll Ggradient for processing poor quality semen samples. Fertil Steril, 1996, 66(2): 459-462.
160. Fulgham DL, Alexander N J, Spermatozoa washing and concentration techniques. In: CRC Handbook of the Laboratory Diagnosis and Treatment of Infertility. Keel BA, webster BW, CRC Press, 1990; 193-212.
161．李宏军．精子体外处理技术．男性学杂志，1994，8(3)：188-190．
162．朱伟杰，刘学高．人类冷冻精液的解冻后处理研究．中国病理生理杂志，1994，10(专刊)：154．
163．营英强，江赛男，董协良．人工授精和传染病．生殖与避孕，1990，10(2)：73-75．
164. Eliasson R. Analysis of semen. In: The Testis. Burger H, ole kretser D, eds, Raven Press, 1981: 381-398.
165. Shanis BS, Check JH, Baker AF. Transmission of Sexually transmitted diseases by donor semen. Arch Androl, 1989, 23(3): 249-257.
166. Greenblatt RM, Handsfield HH, Sayers MH. Screening therapeutic insemination donors for sexually transmitted diseases: overview and recommendation. Fertil Steril, 1986, 43(3): 351-364.
167. Prins GS, Weidel L. A Comparative study of buffer systems as cryoprotectants for human spermatozoa. Fentil Steril, 1986, 46(1): 146-148.168. Weidel L, Prins GS. Cryosurvival of human spermatozoa frozen in eight different buffer systems. J Androl, 1987, 8(1):41-47.
169. Kovacs G, Baker G, Burger H, et al. Artificial insemination with cryopreserved donor semen: a decade of experience. British J Obstet Gynaecol, 1988, 95(3):354-360.
170. Wong AWY, Kan M, Ma HK. Factors affecting the success of artificial insemination by frozen donor semen. Int. J Fertil, 1989, 34(1 ):25-29.
171. Aiman J. Factors affecting the success of donor insemination. Fertil Steril, 1982, 37(1): 94-99.
172. Lamprecht IA, Kotzur B, Schopf E. Round-headed human sperm. Fentil Steril, 1976, 27(6): 685-693.
173. Pederson J, Rebbe H. Fine structure of round-headed human spermatozoa. J Reprod Fertil, 1974, 37(1):51-54.
174. Flork-Gerloff SF, Toper-Peterson E, Muller-Ester. W, et al. Biochemical and genetic investigation of round-headed spermatozoa in infertile men induding two brother and their father. Androlozia, 1984, 16(2):187-202.
175. Check JH, Kozak J, Katsoff D. Hypoosmotic swelling: evalnation of sperm tails using stict criteria in cases of globozoospermia. Arch Androl, 1993, 30(2): 125-128.
176. Lalonde L, Langlais J, Antakl P, et al. Male infertility associated with round-headed acrosomeless spermatozoa. Fertil Steril, 1988, 49(2):316-321.
177. Weissenberg R, Eshkol A, Rudak E, et al. Inability of round-headed acrosomeless human spermatozoa to penetrate zona-free hamster ova. Arch Androl, 1982, 11 (2): 167-169.
178. Syms AJ, Johnson AR, Lip Shultz, LI, et al. Studies on human spermatozoa with round head syndrome. Andrologia, 1984, 42(4):431-435.
179. Rybouchkin AV, Sutter PD, Straeten FV, et al. Fertilization and pregnancy after assisted oocyte activation and intracytoplasmic sperm injection in a case of round-headed sperm associated with deficient oocyte activation capacity. Fertil Steril, 1997, 68(6): 1144-1147.
180. Battaglia DE, Koebler JK, Klein NA, et al. Failure ofoocyte activation after intracytoplasmic sperm injection using round-headed sperm. Fertil Steril, 1997, 68(1): 118-122.
181. Bourne H, Liu DY, Clarke GN, et al. Normal fertilization and embryo development by intracytoplasmic sperm injection of round-headed acrosomeless sperm. Fertil Steril, 1995, 63(6): 1329-1332.
182. Cha K Y, Oum KB, Kim HJ. Approaches for obtaining sperm in patients with male factor infertility, Fertil Steril, 1997, 67(6): 985-995.
183．朱伟杰，刘学高．精子免疫与免疫不育。见：遗传优生与生殖工程．梁志成，朱嘉铭，朱伟杰等编．暨南大学出版社，1992：211-216．
184. Bronson R, Cooper G, Rosenfeld. Sperm antibdies: their role in infertility. Fertil Steril, 1984, 42(2): 171-183.
185. Geva E, Amit A, Lerner-Geval L, et al. Autoimmunity and reproduction. Fertil Steril, 1997, 67(4): 599-611.
186. Kamada M, Yamano S, Senuma M, et al. Semen analysis and antisperm antibody. Arch Androi, 1998, 40(2): 117-128.
187. Naz RK, Menge A. Antisperm antibodies: origin, regulation, and sperm reactivity in human infertility. Fertil Steril, 1994, 61(6): 1001-1013.
188．朱伟杰，梁蔚波，会庆骝等．不育患者睾丸活检前、后的血清抗精子抗体检测．见：广东省生物工程学会论文集．刘学高，朱伟杰编，暨南大学出版社，1992：45-47．
189. Alexander NJ. Treatment for antisperm antibodies: voodoo or victory???Fetil Steril. 1990. 53(4): 602-603.
190．刘继红，章咏裳．免疫与男性不育症．见：生殖免疫学．罗丽兰主编，湖北科技出版社，1998：184-198．
191. Hass GG, Cunningham M, Culp L. The effect of freezing on spermassociated immunoglobulin G(IgG). Fertii Steril, 1984, 42(5): 761-764.
192. Vazquez-Levin MH, Notrica JA, Fried EP. Male immunologic infertility: sperm performance on in vitro fertilization. Fertil Stril, 1997, 68(4): 675-681.
193. Clarke GN, Bourne H, Baker HWG. Intracytoplasmic sperm injection for treating infertility associated with sperm antoimmunity. Fertil Steril, 1997, 68(1): 112-117.
194. Marshburn PB, Kuttch, WH. The role of antisperm antibodies in infertility. Fertil Steril, 1994, 61(5): 799-811.