pGM-CSF/SS基因免疫对雌鼠生长和繁殖的影响及其安全性研究
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摘要
本研究应用质粒提取、细菌培养、PCR、酶连免疫吸附试验(ELISA)、放射免疫测定(RIA)等技术,利用本实验室现有的以减毒沙门氏菌(CSO22)呈递的pGM-CSF/SS基因疫苗,口服免疫45只5周龄雌鼠,以空菌和PBS作为对照,研究生长抑素对生长和繁殖机能的影响及其生物学安全性。主要包括以下内容:
1基因免疫对母鼠的促生长效果
在初次免疫后2周、加强免疫后2周和4周,疫苗免疫组母鼠增重同空菌组和PBS对照组的增重相比差异显著(P＜0.05);空菌组的增重高于PBS对照组,但差异不显著(P＞0.05)。这些结果显示,SS疫苗免疫后对母鼠具有良好的促生长效果。
2基因免疫对生长抑素抗体的影响
基因疫苗免疫组在初次免疫后2周、加强免疫后2周和4周、母鼠分娩当天,血中抗体水平均显著高于空菌组和PBS对照组(P＜0.05),特别是在加强免疫第4周,血中抗体水平与对照组比较差异极显著(P＜0.001),但哺乳3周后,疫苗处理组与对照组差异不显著(P＞0.05)。在SS基因免疫组,抗体水平一开始呈现逐步上升的趋势,到加强免疫后4周出现高峰,但分娩后抗体水平又逐渐下降,到哺乳3周后和对照组差异不显著(p＞0.05)。pGM-CSF/SS基因疫苗免疫可引起较高的体液免疫反应,但抗体持续时间有一定限度。
3基因免疫对母鼠繁殖机能的影响
基因免疫后,对母鼠的发情周期、窝产仔数、窝重和初生重等繁殖指标进行检测,结果显示基因免疫组的发情周期比同空菌组和PBS对照组显著缩短(P＜0.05),说明生长抑素疫苗免疫影响母鼠的发情。窝产仔数、窝重和初生重结果显示,疫苗组同空菌组和PBS对照组相比,差异不显著(P＞0.05),但试验组仔鼠的初生重高于空菌组和PBS对照组。这些结果说明,疫苗免疫可缩短母鼠的发情周期,但对窝产仔数、窝重、初生重没有影响。
测定血中雌激素和孕激素水平显示,在分娩当天,基因免疫组雌激素水平显著高于空菌组和PBS对照组(P＜0.05),而在分娩后3周,各组雌激素都有所下降,但各组差异不显著(P＞0.05);同样,但产后和哺乳3周时孕激素水平各组差异不显著。这些结果表明,SS疫苗免疫对雌激素分泌有促进作用,但对孕激素分泌无影响。
4基因免疫对仔鼠生长发育的影响
哺乳期仔鼠增重结果显示,在产后1周和2周,实验组的增重显著高于空菌组和PBS对照组(P＜0.05),但在哺乳末期,其增重差异不显著(P＞0.05)。哺乳期过后,基因免疫组和空菌组、PBS对照组相比,虽各周增重有所提高,但差异不显著(P＞0.05);哺乳期后,同其它两组相比,基因免疫组的增重效果不显著(P＞0.05)。
仔鼠生后第3、6、9周,基因免疫组同空菌组和PBS对照组相比,抗体水平和生长激素水平有所提高,但差异不显著(P＞0.05),结果同增重效果相吻合。这些结果说明,SS疫苗对母鼠的泌乳性能有一定改善,但哺乳期后,促生长效应不明显。
5基因疫苗的生殖毒性和基因整合研究
母鼠窝产仔数、窝重、仔鼠初重、雌雄比例、子代畸形率和死亡率、母鼠和仔鼠的脏器指数结果显示,基因免疫组同空菌组和对照组相比,差异不显著(P＞0.05)。说明SS疫苗不会影响性成熟母鼠的正常生殖及子代的正常生长发育,对后代无致畸、致死的影响;应用PCR和琼脂糖凝胶电泳进行基因随机整合检测,基因免疫组母鼠和仔鼠各组织0.3μg基因组DNA整合质粒远远小于10个拷贝,远远低于基因组自发突变的概率(2×10~(-6)),表明SS疫苗发生整合的可能性很低,构建的pGM-CSF/SS基因疫苗具有较高的生物安全性。
The pGM-CSF/SS was a previously constructed DNA vaccine against somatostatin (SS) in our laboratory. Its effect on growth and reproduction and its biosafety were studied in this paper using 45 female mice aged 5 weeks. Mice were orally administered with pGM-CSF/SS plasmid that delivered by attenuated Salmonella CSO22, attenuated Salmonella,and PBS (control group) respectively. Techniques such as PCR, ELISA, plasmid extraction and RIA were used to detect gene intergration,antibodies against SS, levels of estradiol and progesterone respectively. The results were shown as follows:
1 The effect of pGM-CSF/SS immunization on growth of female mice
The weight gain in immunized groups was found significantly different compared to the other two groups (p<0.05) at week 2 after the primary and the booster immunization and at week 2 after booster immunization. No significant difference was observed in control and vaccine free groups (p>0.05). This indicated pGM-CSF/SS could promote the growth of immunized female mice.
2 Level of antibodies against SS
The antibodies against SS in immunized mice were found significantly different compared to the control groups (p<0.05) at week 2 after the primary and the booster immunization, week 4 after booster immunization and at the time of pregnance. There were not significant difference among SS immunized groups and the other groups after 3 weeks of lactation (P>0.05). The level of antibodies in SS group showed a trendency of increase and reached the peak at week 4 after immunization. But the level of antibodies decreased gradually after parturition, which showed no significant difference with control group 3 weeks after parturition. pGM-CSF/SS gene vaccine can result a high level of humoral immune reaction, which will last for a certain period of time.
3 The effect of pGM-CSF/SS immunization on reproductive characteristics of female mice
The vaccine group had significantly shorter estrous cycle than other two groups. However, the results about litter size, individual birth weight and birth weight per litter showed no significant difference among three groups (P>0.05). These indicated that pGM-CSF/SS had no significant effect on the litter size, individual birth weight and birth weight per litter, except for shorteninging the estrous cycle.
The levels of estradiol and progesterone were examined at the day of partuition. The result showed that the estrogen level was higher in vaccine group than that in other two control groups (p<0.05). Three weeks later, estrogen level decreased in all three groups. And the difference was not significant between the vaccine group and the other two groups. .However, progesterone level in vaccine group was not significant different than in other groups at the day of partuition and three weeks later(P>0.05). These results indicated that pGM-CSF/SS could affect the estrogen level, but have no effect on progesterone.
4 The effect on the growth of the offspring mice
We examined the weight gain of offspring mice during lactation period. The weight gain in vaccine group was significantly higher than that of the other two groups at the end of the first week and the second week after laboring(p<0.05). However, at the end of lactation period, the difference in weight gain in offspring mice was not significant among three groups(P>0.05). These indicated that the vaccine had limited effect on the ability of lactation. After lactation period, there was no significant difference of weight gain among the offspring mice.
We also examine the level of antibodies against SS and growth hormone of offspring mice. Compared to the pGM-CSF/SS free group and the control, the level of antibodies against SS and growth hormone was higher in vaccine group but not significant(P>0.05). This was in agreement to the weight gained .These results showed that pGM-CSF/SS had limited enhancement of growth of offspring mice.
5 studies on reproductive toxicity and gene integration of DNA vaccine
Comparing the experimental groups with control,there were no difference in Utter size、birth weight per litter、individual birth weight、male/feale of offspring and every organ index in female mice and their offsprings(P>0.05),and there was no malformation and death in offspring. SS DNA vaccine immunization had not bad effect on mice reproduction and their offspring's growth and develepment. No plasmid was detected in all the tissues of mother mice and their musculus, and no integration was found in the investigated tissues. These results indicated that the pGM-CSF/SS delivered by attenuated Salmonella was biosafe.
1基因免疫对母鼠的促生长效果
在初次免疫后2周、加强免疫后2周和4周,疫苗免疫组母鼠增重同空菌组和PBS对照组的增重相比差异显著(P＜0.05);空菌组的增重高于PBS对照组,但差异不显著(P＞0.05)。这些结果显示,SS疫苗免疫后对母鼠具有良好的促生长效果。
2基因免疫对生长抑素抗体的影响
基因疫苗免疫组在初次免疫后2周、加强免疫后2周和4周、母鼠分娩当天,血中抗体水平均显著高于空菌组和PBS对照组(P＜0.05),特别是在加强免疫第4周,血中抗体水平与对照组比较差异极显著(P＜0.001),但哺乳3周后,疫苗处理组与对照组差异不显著(P＞0.05)。在SS基因免疫组,抗体水平一开始呈现逐步上升的趋势,到加强免疫后4周出现高峰,但分娩后抗体水平又逐渐下降,到哺乳3周后和对照组差异不显著(p＞0.05)。pGM-CSF/SS基因疫苗免疫可引起较高的体液免疫反应,但抗体持续时间有一定限度。
3基因免疫对母鼠繁殖机能的影响
基因免疫后,对母鼠的发情周期、窝产仔数、窝重和初生重等繁殖指标进行检测,结果显示基因免疫组的发情周期比同空菌组和PBS对照组显著缩短(P＜0.05),说明生长抑素疫苗免疫影响母鼠的发情。窝产仔数、窝重和初生重结果显示,疫苗组同空菌组和PBS对照组相比,差异不显著(P＞0.05),但试验组仔鼠的初生重高于空菌组和PBS对照组。这些结果说明,疫苗免疫可缩短母鼠的发情周期,但对窝产仔数、窝重、初生重没有影响。
测定血中雌激素和孕激素水平显示,在分娩当天,基因免疫组雌激素水平显著高于空菌组和PBS对照组(P＜0.05),而在分娩后3周,各组雌激素都有所下降,但各组差异不显著(P＞0.05);同样,但产后和哺乳3周时孕激素水平各组差异不显著。这些结果表明,SS疫苗免疫对雌激素分泌有促进作用,但对孕激素分泌无影响。
4基因免疫对仔鼠生长发育的影响
哺乳期仔鼠增重结果显示,在产后1周和2周,实验组的增重显著高于空菌组和PBS对照组(P＜0.05),但在哺乳末期,其增重差异不显著(P＞0.05)。哺乳期过后,基因免疫组和空菌组、PBS对照组相比,虽各周增重有所提高,但差异不显著(P＞0.05);哺乳期后,同其它两组相比,基因免疫组的增重效果不显著(P＞0.05)。
仔鼠生后第3、6、9周,基因免疫组同空菌组和PBS对照组相比,抗体水平和生长激素水平有所提高,但差异不显著(P＞0.05),结果同增重效果相吻合。这些结果说明,SS疫苗对母鼠的泌乳性能有一定改善,但哺乳期后,促生长效应不明显。
5基因疫苗的生殖毒性和基因整合研究
母鼠窝产仔数、窝重、仔鼠初重、雌雄比例、子代畸形率和死亡率、母鼠和仔鼠的脏器指数结果显示,基因免疫组同空菌组和对照组相比,差异不显著(P＞0.05)。说明SS疫苗不会影响性成熟母鼠的正常生殖及子代的正常生长发育,对后代无致畸、致死的影响;应用PCR和琼脂糖凝胶电泳进行基因随机整合检测,基因免疫组母鼠和仔鼠各组织0.3μg基因组DNA整合质粒远远小于10个拷贝,远远低于基因组自发突变的概率(2×10~(-6)),表明SS疫苗发生整合的可能性很低,构建的pGM-CSF/SS基因疫苗具有较高的生物安全性。
The pGM-CSF/SS was a previously constructed DNA vaccine against somatostatin (SS) in our laboratory. Its effect on growth and reproduction and its biosafety were studied in this paper using 45 female mice aged 5 weeks. Mice were orally administered with pGM-CSF/SS plasmid that delivered by attenuated Salmonella CSO22, attenuated Salmonella,and PBS (control group) respectively. Techniques such as PCR, ELISA, plasmid extraction and RIA were used to detect gene intergration,antibodies against SS, levels of estradiol and progesterone respectively. The results were shown as follows:
1 The effect of pGM-CSF/SS immunization on growth of female mice
The weight gain in immunized groups was found significantly different compared to the other two groups (p<0.05) at week 2 after the primary and the booster immunization and at week 2 after booster immunization. No significant difference was observed in control and vaccine free groups (p>0.05). This indicated pGM-CSF/SS could promote the growth of immunized female mice.
2 Level of antibodies against SS
The antibodies against SS in immunized mice were found significantly different compared to the control groups (p<0.05) at week 2 after the primary and the booster immunization, week 4 after booster immunization and at the time of pregnance. There were not significant difference among SS immunized groups and the other groups after 3 weeks of lactation (P>0.05). The level of antibodies in SS group showed a trendency of increase and reached the peak at week 4 after immunization. But the level of antibodies decreased gradually after parturition, which showed no significant difference with control group 3 weeks after parturition. pGM-CSF/SS gene vaccine can result a high level of humoral immune reaction, which will last for a certain period of time.
3 The effect of pGM-CSF/SS immunization on reproductive characteristics of female mice
The vaccine group had significantly shorter estrous cycle than other two groups. However, the results about litter size, individual birth weight and birth weight per litter showed no significant difference among three groups (P>0.05). These indicated that pGM-CSF/SS had no significant effect on the litter size, individual birth weight and birth weight per litter, except for shorteninging the estrous cycle.
The levels of estradiol and progesterone were examined at the day of partuition. The result showed that the estrogen level was higher in vaccine group than that in other two control groups (p<0.05). Three weeks later, estrogen level decreased in all three groups. And the difference was not significant between the vaccine group and the other two groups. .However, progesterone level in vaccine group was not significant different than in other groups at the day of partuition and three weeks later(P>0.05). These results indicated that pGM-CSF/SS could affect the estrogen level, but have no effect on progesterone.
4 The effect on the growth of the offspring mice
We examined the weight gain of offspring mice during lactation period. The weight gain in vaccine group was significantly higher than that of the other two groups at the end of the first week and the second week after laboring(p<0.05). However, at the end of lactation period, the difference in weight gain in offspring mice was not significant among three groups(P>0.05). These indicated that the vaccine had limited effect on the ability of lactation. After lactation period, there was no significant difference of weight gain among the offspring mice.
We also examine the level of antibodies against SS and growth hormone of offspring mice. Compared to the pGM-CSF/SS free group and the control, the level of antibodies against SS and growth hormone was higher in vaccine group but not significant(P>0.05). This was in agreement to the weight gained .These results showed that pGM-CSF/SS had limited enhancement of growth of offspring mice.
5 studies on reproductive toxicity and gene integration of DNA vaccine
Comparing the experimental groups with control,there were no difference in Utter size、birth weight per litter、individual birth weight、male/feale of offspring and every organ index in female mice and their offsprings(P>0.05),and there was no malformation and death in offspring. SS DNA vaccine immunization had not bad effect on mice reproduction and their offspring's growth and develepment. No plasmid was detected in all the tissues of mother mice and their musculus, and no integration was found in the investigated tissues. These results indicated that the pGM-CSF/SS delivered by attenuated Salmonella was biosafe.
引文
1.曹少先,杨利国,茆达干,张文伟,管峰.生长抑素基因疫苗pcS/SS构建及其在HeLa细胞中的表达.中国兽医学报,2004,24:153-156
2.曹少先,张文伟,茆达干,管峰,杨利国.生长抑素基因疫苗质粒pcS/2SS的构建、表达及免疫.农业生物技术学报,2005a,13(4):477-481
3.曹少先,杨利国,张文伟,茆达干,何晓红.生长抑素基因疫苗质粒pEGS/2SS 的构建及表达.中国兽医学报,2005b,25:499-502
4.曹少先,何晓红,孙延鸣,刘铁铮,杨利国.生长抑素基因疫苗pES/2SS的构建与表达.扬州大学学报(农业与生命科学版),2007,28:5-8
5.程从升,王文成,李素,扈荣良,涂长春,刘江徐.猪瘟dna疫苗在猪体及环境的生物安全性研究.微生物学报,2005,45(2):292-297
6.程丹,王国文.药物基因组学及生物芯片应用.实用药物与临床,2008,11(02):114-115
7.付洁,宋海峰.核酸疫苗的药动学与安全性研究进展.中国新药杂志,2006,15(5):337-340
8.高学勇,陈奕权,何为民.小鼠生殖周期子宫、血生长抑素的变化.福建医科大学学报,2000,34:24-26
9.何晓红.生长抑素和卵泡抑制素基因重组质粒转染对猪和牛卵泡颗粒细胞的作用.[硕士学位论文].南京:南京农业大学图书馆,2004
10.胡斌,莫国玉,刘燕,蔡颖鹏,程钢,何蕴韶.荧光定量PCR技术检测DNA 疫苗在生殖腺和血液中的残留.热带医学杂志,2006,6:991-996
11.李雪梅,朱文杰,陈秀敏.生长激素在体外受精周期中的应用初探.实用医学杂志,2004,20:883-884
12.杨利国主编,动物繁殖学.北京:中国农业出版社,2002,366-367
13.梁爱心,姚艳丰,金定恩,武睿,韩丽,付平,刘耘,杨利国.小鼠对pGM-CSF/SS 生长抑素基因疫苗的免疫应答.华中农业大学学报,2006,25(5):540-543
14.梁昊宇,曾明.新疫苗和佐剂的临床前安全性评价.国际生物制品学杂志,2006,29:202-205
15.舒邓群,茆达干,吴志敏,程宝,杨利国.以减毒沙门氏菌为载体的Gm-Csf 与生长抑素融合表达质粒对小鼠淋巴细胞增殖和gh及Igf-Ⅰ分泌的影响.畜牧兽医学报,2006a,37(8):814-818
16.舒邓群.核酸佐剂GM-CSF的克隆及其对生长抑素DNA疫苗的免疫增强作用.[博士学位论文].南京:南京农业大学图书馆,2006b
17.舒邓群,茆达干,陈荣达,吴志敏,杨利国.粒细胞-巨噬细胞集落刺激因子与 生长抑素融合表达质粒对断奶仔猪生长及相关激素的影响.南京农业大学学报,2007,30(4):92-96
18.孙伟,崔薇,闻姬.两种不同剂量生长激素在体外受精超排卵中的作用比较.中国优生与遗传杂志,2002,10:9-10
19.王颖,李美芝,杨池荪,王振红,姜颜彬.生长激素在多囊卵巢综合征促排卵中的作用.中华妇产科杂志,2001,36:36-39
20.吴琳,刁振宇,邓小昭,高健,周宗安,刘玉,王元伦.猪囊虫Dna疫苗在猪体内的组织分布及其安全性探讨.药物生物技术,2006,13(1):14-16
21.向丽华,陈燕萍,张智,闪增郁,于智敏,吕爱平.24味有毒中药长期毒性实验对大鼠脏器指数的影响.中国中医基础医学杂志,2006,1:35-37
22.徐文忠,陈萍.免疫中和体内生长抑素促进动物生长.国外畜牧科技,1992,19(2):17-19
23.徐文忠,杜念兴,李光地.生长抑素与乙肝表面抗原融合基因重组痘苗病毒的构建及其表达[J].南京农业大学学报,1992,15(3):74-80
24.薛春林,茆达干,杨利国,程宝.基因佐剂CpGDNA对生长抑素DNA疫苗pES/2SS的作用.中国农业科学,2007a,6(7):101-105
25.薛春林,茆达干,杨利国,王勇.生长抑素dna疫苗对湖羊羔羊生长和相关激素的作用——佐剂效应.南京农业大学学报,2007b,30(2):89-93
26.姚艳丰.一种生长抑素基因免疫的新策略研究.[硕士学位论文].武汉:华中农业大学图书馆,2007
27.杨锦波,刘天佳,李继遥.抗龋基因疫苗pcDNA3-gtfB整合宿主细胞基因组DNA 的可能性研究.华西口腔医学杂志,2003,21(3):228-230
28.郑风荣,孙修勤,刘洪展,吴谡琦,张进兴,曲凌云,洪旭光.牙鲆淋巴囊肿病毒核酸疫苗的安全性研究.高技术通讯,2006,16(1):106-110
29.曾义祥,杜念兴.生长抑制激素基因在大肠杆菌中的表达.遗传学报,1991,18(3):282-288
30.Aixin Liang,Shaoxian Cao,LiHan,Yanfeng Yao,M.Moaeen-ud-Din,Liguo Yang.Construction and evaluation of the eukaryotic expression plasmid encoding two copies of soatostatin genes fused with hepatitis B surface antigen gene S.Vaccine,2008,26:2935-2941
31.Andreani CL,Lazzarin N,Pierro E,Lanzone A,Mancuso S.Somatostatin action on rat ovarian steroidogenesis.Hum Reprod,1995,10(8):1968-73
32.Ashok MS,Rangarajan PN.Protective efficacy of a plasmid DNA encoding Japanese encephalitis virus envelope protein fused to tissue plasminogen activator signal sequences:studies in a murine intracerebral virus challenge model.Vaccine,2002, 20(11-12):1563-70
33. Aurich JE, Kranski S, Parvizi N, Aurich C. Somatostatin treatment affects testicular function in stallions. Theriogenology, 2003, 60(1): 163-74
34. Bartke A, Chandrashekar V, Turyn D, Steger RW, Debeljuk L,WintersTA, Mattison JA, Danilovich NA, Croson W, Wernsing DR, KopchickJJ. Effects of growth hormone overexpression and growth hormoneresistance on neuroendocrine and reproductive functions in transgenicand knock-out mice. Proc Soc Exp Biol Med, 1999,222:113-123
35. Bartke A, Chandrashekar V, Bailey B, Zaczek D, Turyn D. Conse quences of growth hormone (GH) overexpression and GH resistance. Neuropeptides, 2002,36:201-208
36. Brennan FR, Dougan G. Non-clinical safety evaluation of novel vaccines and adjuvants: new products, new strategies. Vaccine, 2005, 23(24):3210-22
37. Bryan KA, Hagen DR, Hammond JM. Effect of frequency of administration of exogenous porcine growth hormone on growth and carcass traits and ovarian function of prepubertal gilts. J Anim Sci, 1992, 70(5): 1454-63
38. Charland S, Boucher MJ, Houde M, Rivard N. Somatostatin inhibits Akt phosphorylation and cell cycle entry, but not p42/p44 mitogen-activated protein (MAP) kinase activation in normal and tumoral pancreatic acinar cells. Endocrinology, 2001, 142(1): 121-8
39. Chayvialle J,Paulin C,Dubois PM,Descos F,Dubious Mp.Ontcgeny of somatostation in the huan gastrointestinal tract endocrine pancreas and hypothalamus. Acta Endocrinol, 1980,94:1-10
40. Coelho-Castelo AA, Trombone AP, Rosada RS, Santos RR, Jr., Bonato VL, Sartori A, Silva CL. Tissue distribution of a plasmid DNA encoding Hsp65 gene is dependent on the dose administered through intramuscular delivery. Genet Vaccines Ther, 2006,4:1
41. Conti M. Specificity of the cyclic adenosine 3',5'-monophosphate signal in granulosa cell function. Biol Reprod, 2002, 67(6): 1653-61
42. Dietrich G, Spreng S, Gentschev I, Goebel W. Bacterial systems for the delivery of eukaryotic antigen expression vectors. Antisense Nucleic Acid Drug Dev, 2000, 10(5):391-9
43. Ebert M, Friess H, Beger HG, Buchler MW. Role of octreotide in the treatment of pancreatic cancer. Digestion, 1994, 55 Suppl 1:48-51
44. Eon-Duval A, Burke G. Purification of pharmaceutical-grade plasmid DNA by anion-exchange chromatography in an RNase-free process. J Chromatogr B Analyt. Technol Biomed Life Sci, 2004, 804(2):327-35
45. Galton, D. M., R. W. Everett, M. E. Van Amburg, D. L. Bauman,and W. A. Knoblauch.. Extended calving intervals with theuse of bST. Pages in Proc. Western Large Herd Dairy, 1997, 116-121
46. Gerdts V, Babiuk LA, van Drunen Littel-van den H, Griebel PJ. Fetal immunization by a DNA vaccine delivered into the oral cavity. Nat Med, 2000, 6(8):929-32
47. Gerendai I, Csaba Z, Csernus V. Effect of intratesticular administration of somatostatin on testicular function in immature and adult rats. Life Sci, 1996, 59(10):859-66
48. Giannetti N, Enjalbert A, Krantic S. Somatostatin analog SMS 201995 inhibits proliferation in human leukemia T-cell line: relevance of the adenylyl cyclase stimulation. J Cell Biochem, 2000, 78(4):666-673
49. Glenting J, Madsen SM, Vrang A, Fomsgaard A, Israelsen H. A plasmid selection system in Lactococcus lactis and its use for gene expression in L. lactis and human kidney fibroblasts. Appl Environ Microbiol, 2002, 68(10):5051-5056
50. Haworth R, Pilling AM. The PCR assay in the preclinical safety evaluation of nucleic acid medicines. Hum Exp Toxicol, 2000, 19(5):267-76
51. Hoist N, Haug E, Tanbo T, Abyholm T, Jacobsen MB. Somatostatin in human follicular fluid. Hum Reprod, 1994, 9(8):1448-1451
52. Hoist N, Kierulf KH, Seppala M, Koistinen R, Jacobsen MB. Regulation of insulin-like growth factor-binding protein-1 and progesterone secretion from human granulosa-luteal cells: effects of octreotide and insulin. Fertil Steril, 1997, 68(3):478-82
53. Huang DB, Mohanty A, DuPont HL, Okhuysen PC, Chiang T. A review of an emerging enteric pathogen: enteroaggregative Escherichia coli. J Med Microbiol, 2006, 55(Pt 10):1303-11
54. Hubbard K, Catalano J, Puri RK, Gnatt A. Knockdown of TFIIS by RNA silencing inhibits cancer cell proliferation and induces apoptosis. BMC Cancer, 2008, 8(1):133
55. Kanellos T, Sylvester ID, Ambali AG, Howard CR, Russell PH. The safety and longevity of DNA vaccines for fish. Immunology, 1999, 96(2):307-13
56. Kennedy HE, McCullough SJ, Graham D, Cassidy J, Malone FE, Ellis WA. Detection of chlamydial antibody by fetal serology—an aid to the diagnosis of ovine abortion. J Vet Diagn Invest, 2001, 13(1):30-5
57. Kirkwood RN, Korchinski RS, Thacker PA, Laarveld B. Observations on the influence of active immunization against somatostatin on the reproductive performance of sheep and pigs. J Reprod Immunol, 1990, 17(3):229-38
58. Kirkwood RN, Thacker PA, Korchinski RS, Laarveld B. The influence of exogenous pituitary growth hormone on the ovulatory response to PMSG and hCG and the LH response to estradiol benzoate in prepubertal gilts. Domest Anim Endocrinol, 1988, 5(4):317-22
59. Kitikoon P, Strait EL, Thacker EL. The antibody responses to swine influenza virus (SIV) recombinant matrix 1 (rM1), matrix 2 (M2), and hemagglutinin (HA) proteins in pigs with different SIV exposure. Vet Microbiol, 2008, 126(1-3):51-62
60. Klinman DM, Takeshita F, Kamstrup S, Takeshita S, Ishii K, Ichino M, Yamada H. DNA vaccines: capacity to induce auto-immunity and tolerance. Dev Biol (Basel), 2000, 104:45-51
61. Kouroumalis E, Skordilis P, Thermos K, Vasilaki A, Moschandrea J, Manousos ON. Treatment of hepatocellular carcinoma with octreotide: a randomised controlled study. Gut, 1998, 42(3):442-7
62. Kouroumalis EA. Octreotide for cancer of the liver and biliary tree. Chemotherapy, 2001, 47 Suppl 2:150-61
63. Krieg AM, Yi AK, Matson S, Waldschmidt TJ, Bishop GA, Teasdale R, Koretzky GA, Klinman DM. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature, 1995, 374(6522):546-9
64. Lachowicz-Ochedalska A, Rebas E, Kunert-Radek J, Winczyk K, Pawlikowski M. Effects of somatostatin and its analogues on tyrosine kinase activity in rodent tumors. Biol Signals Recept, 2000, 9(5):255-9
65. Lebron JA, Wolf JJ, Kaplanski CV, Ledwith BJ. Ensuring the quality, potency and safety of vaccines during preclinical development. Expert Rev Vaccines, 2005, 4(6):855-66
66. Ledwith BJ, Manam S, Troilo PJ, Barnum AB, Pauley CJ, Griffiths TG, 2nd, Harper LB, Beare CM, Bagdon WJ, Nichols WW. Plasmid DNA vaccines: investigation of integration into host cellular DNA following intramuscular injection in mice. Intervirology, 2000, 43(4-6):258-72
67. LiHan,D.G..Mao,D.K.Zhang,A.X.Liang,M.Fang,MuhamadMoaeen-ud-Din,L.G.Yang Development and evaluation of a novel DNA vaccine expressing inhibin (1-32) fragment for improving the fertility in rats and sheep. Animal Repr Sci, 2008
68. Liu JW, Cheng J. Molecular mechanism of immune response induced by foreign plasmid DNA after oral administration in mice. World J Gastroenterol, 2007, 13(28):3847-54
69. Luna-Dominguez JE, Enns RM, Armstrong DV, Ax RL. Reproductive performance of Holstein cows receiving somatotropin. J Dairy Sci, 2000, 83(7): 1451-5
70. Manam S, Ledwith BJ, Barnum AB, Troilo PJ, Pauley CJ, Harper LB, Griffiths TG, 2nd, Niu Z, Denisova L, Follmer TT and others. Plasmid DNA vaccines: tissue distribution and effects of DNA sequence, adjuvants and delivery method on integration into host DNA. Intervirology, 2000, 43(4-6):273-81
71. McIntyre HD, Marechal DJ, Deby GP, Mathieu AG, Hezee-Hagelstein MT, Franchimont PP. Immunoreactive somatostatin in the rat ovary. Acta Endocrinol (Copenh), 1992, 126(6):553-8
72. Mor G, Yamshchikov G, Sedegah M, Takeno M, Wang R, Houghten RA, Hoffman S, Klinman DM. Induction of neonatal tolerance by plasmid DNA vaccination of mice. J Clin Invest, 1996, 98(12):2700-5
73. Mori T, Saito H, Ohno Y, Irahara M, Hosoi E, Saito S. Evidence for existence of somatostatin-like immunoreactivity with molecular heterogeneity in porcine ovaries. Acta Endocrinol (Copenh), 1984, 106(2):254-9
74. Naar EM, Bartke A, Majumdar SS, Buonomo FC, Yun JS, WagnerTE. Fertility of transgenic female mice expressing bovine growth hormoneor human growth hormone variant genes. Biol Reprod, 1991, 45:178-187
75. Nishiyama K, Sugimoto T, Kaji H, Kanatani M, Kobayashi T, Chihara K. Stimulatory effect of growth hormone on bone resorption and osteoclast differentiation. Endocrinology, 1996, 137(1):35-41
76. Owens PC, Conlon MA, Campbell RG, Johnson RJ, King R, Ballard FJ. Developmental changes in growth hormone, insulin-like growth factors (IGF-I and IGF-II) and IGF-binding proteins in plasma of young growing pigs. J Endocrinol, 1991, 128(3):439-47
77. Parker SE, Borellini F, Wenk ML, Hobart P, Hoffman SL, Hedstrom R, Le T, Norman JA. Plasmid DNA malaria vaccine: tissue distribution and safety studies in mice and rabbits. Hum Gene Ther, 1999, 10(5):741-58
78. Piedra PA. Clinical experience with respiratory syncytial virus vaccines. Pediatr Infect Dis J, 2003, 22(2 Suppl):S94-9
79. Pilling AM, Harman RM, Jones SA, McCormack NA, Lavender D, Haworth R. The assessment of local tolerance, acute toxicity, and DNA biodistribution following particle-mediated delivery of a DNA vaccine to minipigs. Toxicol Pathol, 2002, 30(3):298-305
80. Pillon D, Caraty A, Fabre-Nvs C, Lomet D, Cateau M, Bruneau G. Regulation by estradiol of hypothalamic somatostatin gene expression: possible involvement of somatostatin in the control of luteinizing hormone secretion in the ewe.Biol Reprod, 2004, 71(1):38-44
81. Pisetsky DS. Immune response to DNA in systemic lupus erythematosus. Isr Med Assoc J, 2001, 3(11):850-3
82. Pollak MN, Schally AV. Mechanisms of antineoplastic action of somatostatin analogs. Proc Soc Exp Biol Med, 1998, 217(2): 143-52
83. Rajkumar K, Kerr DE, Kirkwood RN, Laarveld B. Inhibitory action of somatostatin-14 on hormone-stimulated cyclic adenosine monophosphate induction in porcine granulosa and luteal cells. J Endocrinol, 1992, 134(2):297-306
84. Schalk JA, Mooi FR, Berbers GA, van Aerts LA, Ovelgonne H, Kimman TG. Preclinical and clinical safety studies on DNA vaccines. Hum Vaccin, 2006, 2(2):45-53
85. Schneider G, Fechner U. Advances in the prediction of protein targeting signals. Proteomics, 2004, 4(6): 1571-80
86. Shata MT, Stevceva L, Agwale S, Lewis GK, Hone DM. Recent advances with recombinant .bacterial vaccine vectors.Mol Med Today, 2000, 6(2):66-71
87. Sheets RL, Stein J, Manetz TS, Andrews C, Bailer R, Rathmann J, Gomez PL. Toxicological safety evaluation of DNA plasmid vaccines against HIV-1, Ebola, Severe Acute Respiratory Syndrome, or West Nile virus is similar despite differing plasmid backbones or gene-inserts. Toxicol Sci, 2006, 91(2):620-30
88. Shiau AL, Chu CY, Su WC, Wu CL. Vaccination with the glycoprotein D gene of pseudorabies virus delivered by nonpathogenic Escherichia coli elicits protective immune responses. Vaccine, 2001, 19(23-24):3277-84
89. Spencer GS, Hallett KG, Seddon A, Morris BA. Immuno-neutralization of maternal plasma oestradiol stimulates fetal growth in rats. Reprod Nutr Dev, 1986, 26(6): 1289-93
90. Spreng S, Viret JF. Plasmid maintenance systems suitable for GMO-based bacterial vaccines. Vaccine, 2005, 23(17-18):2060-5
91. Thomas AD, Murray JD, Famula TR, Oberbauer AM. Growth hormone and fertility in oMt1a-oGH transgenic mice. Reproduction, 2001, 122(4):537-44
92. Tran ND, Porada CD, Almeida-Porada G, Glimp HA, Anderson WF, Zanjani ED. Induction of stable prenatal tolerance to beta-galactosidase by in utero gene transfer into preimmune sheep fetuses. Blood, 2001, 97(11):3417-23
93. Tsang CH, Mirakhur KK, Babiuk LA, Griebel PJ. Oral DNA immunization in the second trimester fetal lamb and secondary immune responses in the neonate. Vaccine, 2007, 25(50):8469-79
94. Tseng J C, Levin B, Hurtado A, Yee H, Perez de Castro I,Jimenez M,Shamamian P,JinR,Novick RP,Pellicer A Meruelo D . Systemic tumor targeting and killing by sindbis viral vectors. NatBiotechnol, 2004, 22:70-77
95. Vernejoul F, Faure P, Benali N, Calise D, Tiraby G, Pradayrol L, Susini C, Buscail L. Antitumor effect of in vivo somatostatin receptor subtype 2 gene transfer in primary and metastatic pancreatic cancer models. Cancer Res, 2002, 62(21):6124-31
96. Welle S, Bhatt K, Pinkert CA, Tawil R, Thornton CA. Muscle growth after postdevelopmental myostatin gene knockout. Am J Physiol Endocrinol Metab, 2007, 292(4):E985-91
97. Wolf JJ, Wang L, Wang F. Application of PCR technology in vaccine product development. Expert Rev Vaccines, 2007,6(4):547-58
98. Wraith DC, Goldman M, Lambert PH. Vaccination and autoimmune disease: what is the evidence? Lancet, 2003, 362(9396): 1659-66
99. Xing Y, Wensheng L, Haoran L. Polygenic expression of somatostatin in orange-spotted grouper (Epinephelus coioides): molecular cloning and distribution of the mRNAs encoding three somatostatin precursors. Mol Cell Endocrinol, 2005, 241(1-2):62-72
100.Yoon HA, Aleyas AG, George JA, Park SO, Han YW, Lee JH, Cho JG, Eo SK. Cytokine GM-CSF genetic adjuvant facilitates prophylactic DNA vaccine against pseudorabies virus through enhanced immune responses. Microbiol Immunol, 2006, 50(2):83-92
101.Zaczek D, Hammond J, Suen L, Wandji S, Service D, Bartke A, Chandrashekar V, Coschigano K, Kopchick J. Impact of growth hormone resistance on female reproductive function: new insights from growth hormone receptor knockout mice. Biol Reprod, 2002, 67(4): 1115-24
102.Z.Roth, A.Arav, R.Braw-Tal, A.Bor,D.Wolfenson. Effect of Treament with Follicle-Stimulating Hormone or Bovine Somatotropin on the Quality of Oocytes Aspirated in the Autumn from Previously Heat-Stressed Cows. J Dairy Sci, 2002, 85:1398-1405
2.曹少先,张文伟,茆达干,管峰,杨利国.生长抑素基因疫苗质粒pcS/2SS的构建、表达及免疫.农业生物技术学报,2005a,13(4):477-481
3.曹少先,杨利国,张文伟,茆达干,何晓红.生长抑素基因疫苗质粒pEGS/2SS 的构建及表达.中国兽医学报,2005b,25:499-502
4.曹少先,何晓红,孙延鸣,刘铁铮,杨利国.生长抑素基因疫苗pES/2SS的构建与表达.扬州大学学报(农业与生命科学版),2007,28:5-8
5.程从升,王文成,李素,扈荣良,涂长春,刘江徐.猪瘟dna疫苗在猪体及环境的生物安全性研究.微生物学报,2005,45(2):292-297
6.程丹,王国文.药物基因组学及生物芯片应用.实用药物与临床,2008,11(02):114-115
7.付洁,宋海峰.核酸疫苗的药动学与安全性研究进展.中国新药杂志,2006,15(5):337-340
8.高学勇,陈奕权,何为民.小鼠生殖周期子宫、血生长抑素的变化.福建医科大学学报,2000,34:24-26
9.何晓红.生长抑素和卵泡抑制素基因重组质粒转染对猪和牛卵泡颗粒细胞的作用.[硕士学位论文].南京:南京农业大学图书馆,2004
10.胡斌,莫国玉,刘燕,蔡颖鹏,程钢,何蕴韶.荧光定量PCR技术检测DNA 疫苗在生殖腺和血液中的残留.热带医学杂志,2006,6:991-996
11.李雪梅,朱文杰,陈秀敏.生长激素在体外受精周期中的应用初探.实用医学杂志,2004,20:883-884
12.杨利国主编,动物繁殖学.北京:中国农业出版社,2002,366-367
13.梁爱心,姚艳丰,金定恩,武睿,韩丽,付平,刘耘,杨利国.小鼠对pGM-CSF/SS 生长抑素基因疫苗的免疫应答.华中农业大学学报,2006,25(5):540-543
14.梁昊宇,曾明.新疫苗和佐剂的临床前安全性评价.国际生物制品学杂志,2006,29:202-205
15.舒邓群,茆达干,吴志敏,程宝,杨利国.以减毒沙门氏菌为载体的Gm-Csf 与生长抑素融合表达质粒对小鼠淋巴细胞增殖和gh及Igf-Ⅰ分泌的影响.畜牧兽医学报,2006a,37(8):814-818
16.舒邓群.核酸佐剂GM-CSF的克隆及其对生长抑素DNA疫苗的免疫增强作用.[博士学位论文].南京:南京农业大学图书馆,2006b
17.舒邓群,茆达干,陈荣达,吴志敏,杨利国.粒细胞-巨噬细胞集落刺激因子与 生长抑素融合表达质粒对断奶仔猪生长及相关激素的影响.南京农业大学学报,2007,30(4):92-96
18.孙伟,崔薇,闻姬.两种不同剂量生长激素在体外受精超排卵中的作用比较.中国优生与遗传杂志,2002,10:9-10
19.王颖,李美芝,杨池荪,王振红,姜颜彬.生长激素在多囊卵巢综合征促排卵中的作用.中华妇产科杂志,2001,36:36-39
20.吴琳,刁振宇,邓小昭,高健,周宗安,刘玉,王元伦.猪囊虫Dna疫苗在猪体内的组织分布及其安全性探讨.药物生物技术,2006,13(1):14-16
21.向丽华,陈燕萍,张智,闪增郁,于智敏,吕爱平.24味有毒中药长期毒性实验对大鼠脏器指数的影响.中国中医基础医学杂志,2006,1:35-37
22.徐文忠,陈萍.免疫中和体内生长抑素促进动物生长.国外畜牧科技,1992,19(2):17-19
23.徐文忠,杜念兴,李光地.生长抑素与乙肝表面抗原融合基因重组痘苗病毒的构建及其表达[J].南京农业大学学报,1992,15(3):74-80
24.薛春林,茆达干,杨利国,程宝.基因佐剂CpGDNA对生长抑素DNA疫苗pES/2SS的作用.中国农业科学,2007a,6(7):101-105
25.薛春林,茆达干,杨利国,王勇.生长抑素dna疫苗对湖羊羔羊生长和相关激素的作用——佐剂效应.南京农业大学学报,2007b,30(2):89-93
26.姚艳丰.一种生长抑素基因免疫的新策略研究.[硕士学位论文].武汉:华中农业大学图书馆,2007
27.杨锦波,刘天佳,李继遥.抗龋基因疫苗pcDNA3-gtfB整合宿主细胞基因组DNA 的可能性研究.华西口腔医学杂志,2003,21(3):228-230
28.郑风荣,孙修勤,刘洪展,吴谡琦,张进兴,曲凌云,洪旭光.牙鲆淋巴囊肿病毒核酸疫苗的安全性研究.高技术通讯,2006,16(1):106-110
29.曾义祥,杜念兴.生长抑制激素基因在大肠杆菌中的表达.遗传学报,1991,18(3):282-288
30.Aixin Liang,Shaoxian Cao,LiHan,Yanfeng Yao,M.Moaeen-ud-Din,Liguo Yang.Construction and evaluation of the eukaryotic expression plasmid encoding two copies of soatostatin genes fused with hepatitis B surface antigen gene S.Vaccine,2008,26:2935-2941
31.Andreani CL,Lazzarin N,Pierro E,Lanzone A,Mancuso S.Somatostatin action on rat ovarian steroidogenesis.Hum Reprod,1995,10(8):1968-73
32.Ashok MS,Rangarajan PN.Protective efficacy of a plasmid DNA encoding Japanese encephalitis virus envelope protein fused to tissue plasminogen activator signal sequences:studies in a murine intracerebral virus challenge model.Vaccine,2002, 20(11-12):1563-70
33. Aurich JE, Kranski S, Parvizi N, Aurich C. Somatostatin treatment affects testicular function in stallions. Theriogenology, 2003, 60(1): 163-74
34. Bartke A, Chandrashekar V, Turyn D, Steger RW, Debeljuk L,WintersTA, Mattison JA, Danilovich NA, Croson W, Wernsing DR, KopchickJJ. Effects of growth hormone overexpression and growth hormoneresistance on neuroendocrine and reproductive functions in transgenicand knock-out mice. Proc Soc Exp Biol Med, 1999,222:113-123
35. Bartke A, Chandrashekar V, Bailey B, Zaczek D, Turyn D. Conse quences of growth hormone (GH) overexpression and GH resistance. Neuropeptides, 2002,36:201-208
36. Brennan FR, Dougan G. Non-clinical safety evaluation of novel vaccines and adjuvants: new products, new strategies. Vaccine, 2005, 23(24):3210-22
37. Bryan KA, Hagen DR, Hammond JM. Effect of frequency of administration of exogenous porcine growth hormone on growth and carcass traits and ovarian function of prepubertal gilts. J Anim Sci, 1992, 70(5): 1454-63
38. Charland S, Boucher MJ, Houde M, Rivard N. Somatostatin inhibits Akt phosphorylation and cell cycle entry, but not p42/p44 mitogen-activated protein (MAP) kinase activation in normal and tumoral pancreatic acinar cells. Endocrinology, 2001, 142(1): 121-8
39. Chayvialle J,Paulin C,Dubois PM,Descos F,Dubious Mp.Ontcgeny of somatostation in the huan gastrointestinal tract endocrine pancreas and hypothalamus. Acta Endocrinol, 1980,94:1-10
40. Coelho-Castelo AA, Trombone AP, Rosada RS, Santos RR, Jr., Bonato VL, Sartori A, Silva CL. Tissue distribution of a plasmid DNA encoding Hsp65 gene is dependent on the dose administered through intramuscular delivery. Genet Vaccines Ther, 2006,4:1
41. Conti M. Specificity of the cyclic adenosine 3',5'-monophosphate signal in granulosa cell function. Biol Reprod, 2002, 67(6): 1653-61
42. Dietrich G, Spreng S, Gentschev I, Goebel W. Bacterial systems for the delivery of eukaryotic antigen expression vectors. Antisense Nucleic Acid Drug Dev, 2000, 10(5):391-9
43. Ebert M, Friess H, Beger HG, Buchler MW. Role of octreotide in the treatment of pancreatic cancer. Digestion, 1994, 55 Suppl 1:48-51
44. Eon-Duval A, Burke G. Purification of pharmaceutical-grade plasmid DNA by anion-exchange chromatography in an RNase-free process. J Chromatogr B Analyt. Technol Biomed Life Sci, 2004, 804(2):327-35
45. Galton, D. M., R. W. Everett, M. E. Van Amburg, D. L. Bauman,and W. A. Knoblauch.. Extended calving intervals with theuse of bST. Pages in Proc. Western Large Herd Dairy, 1997, 116-121
46. Gerdts V, Babiuk LA, van Drunen Littel-van den H, Griebel PJ. Fetal immunization by a DNA vaccine delivered into the oral cavity. Nat Med, 2000, 6(8):929-32
47. Gerendai I, Csaba Z, Csernus V. Effect of intratesticular administration of somatostatin on testicular function in immature and adult rats. Life Sci, 1996, 59(10):859-66
48. Giannetti N, Enjalbert A, Krantic S. Somatostatin analog SMS 201995 inhibits proliferation in human leukemia T-cell line: relevance of the adenylyl cyclase stimulation. J Cell Biochem, 2000, 78(4):666-673
49. Glenting J, Madsen SM, Vrang A, Fomsgaard A, Israelsen H. A plasmid selection system in Lactococcus lactis and its use for gene expression in L. lactis and human kidney fibroblasts. Appl Environ Microbiol, 2002, 68(10):5051-5056
50. Haworth R, Pilling AM. The PCR assay in the preclinical safety evaluation of nucleic acid medicines. Hum Exp Toxicol, 2000, 19(5):267-76
51. Hoist N, Haug E, Tanbo T, Abyholm T, Jacobsen MB. Somatostatin in human follicular fluid. Hum Reprod, 1994, 9(8):1448-1451
52. Hoist N, Kierulf KH, Seppala M, Koistinen R, Jacobsen MB. Regulation of insulin-like growth factor-binding protein-1 and progesterone secretion from human granulosa-luteal cells: effects of octreotide and insulin. Fertil Steril, 1997, 68(3):478-82
53. Huang DB, Mohanty A, DuPont HL, Okhuysen PC, Chiang T. A review of an emerging enteric pathogen: enteroaggregative Escherichia coli. J Med Microbiol, 2006, 55(Pt 10):1303-11
54. Hubbard K, Catalano J, Puri RK, Gnatt A. Knockdown of TFIIS by RNA silencing inhibits cancer cell proliferation and induces apoptosis. BMC Cancer, 2008, 8(1):133
55. Kanellos T, Sylvester ID, Ambali AG, Howard CR, Russell PH. The safety and longevity of DNA vaccines for fish. Immunology, 1999, 96(2):307-13
56. Kennedy HE, McCullough SJ, Graham D, Cassidy J, Malone FE, Ellis WA. Detection of chlamydial antibody by fetal serology—an aid to the diagnosis of ovine abortion. J Vet Diagn Invest, 2001, 13(1):30-5
57. Kirkwood RN, Korchinski RS, Thacker PA, Laarveld B. Observations on the influence of active immunization against somatostatin on the reproductive performance of sheep and pigs. J Reprod Immunol, 1990, 17(3):229-38
58. Kirkwood RN, Thacker PA, Korchinski RS, Laarveld B. The influence of exogenous pituitary growth hormone on the ovulatory response to PMSG and hCG and the LH response to estradiol benzoate in prepubertal gilts. Domest Anim Endocrinol, 1988, 5(4):317-22
59. Kitikoon P, Strait EL, Thacker EL. The antibody responses to swine influenza virus (SIV) recombinant matrix 1 (rM1), matrix 2 (M2), and hemagglutinin (HA) proteins in pigs with different SIV exposure. Vet Microbiol, 2008, 126(1-3):51-62
60. Klinman DM, Takeshita F, Kamstrup S, Takeshita S, Ishii K, Ichino M, Yamada H. DNA vaccines: capacity to induce auto-immunity and tolerance. Dev Biol (Basel), 2000, 104:45-51
61. Kouroumalis E, Skordilis P, Thermos K, Vasilaki A, Moschandrea J, Manousos ON. Treatment of hepatocellular carcinoma with octreotide: a randomised controlled study. Gut, 1998, 42(3):442-7
62. Kouroumalis EA. Octreotide for cancer of the liver and biliary tree. Chemotherapy, 2001, 47 Suppl 2:150-61
63. Krieg AM, Yi AK, Matson S, Waldschmidt TJ, Bishop GA, Teasdale R, Koretzky GA, Klinman DM. CpG motifs in bacterial DNA trigger direct B-cell activation. Nature, 1995, 374(6522):546-9
64. Lachowicz-Ochedalska A, Rebas E, Kunert-Radek J, Winczyk K, Pawlikowski M. Effects of somatostatin and its analogues on tyrosine kinase activity in rodent tumors. Biol Signals Recept, 2000, 9(5):255-9
65. Lebron JA, Wolf JJ, Kaplanski CV, Ledwith BJ. Ensuring the quality, potency and safety of vaccines during preclinical development. Expert Rev Vaccines, 2005, 4(6):855-66
66. Ledwith BJ, Manam S, Troilo PJ, Barnum AB, Pauley CJ, Griffiths TG, 2nd, Harper LB, Beare CM, Bagdon WJ, Nichols WW. Plasmid DNA vaccines: investigation of integration into host cellular DNA following intramuscular injection in mice. Intervirology, 2000, 43(4-6):258-72
67. LiHan,D.G..Mao,D.K.Zhang,A.X.Liang,M.Fang,MuhamadMoaeen-ud-Din,L.G.Yang Development and evaluation of a novel DNA vaccine expressing inhibin (1-32) fragment for improving the fertility in rats and sheep. Animal Repr Sci, 2008
68. Liu JW, Cheng J. Molecular mechanism of immune response induced by foreign plasmid DNA after oral administration in mice. World J Gastroenterol, 2007, 13(28):3847-54
69. Luna-Dominguez JE, Enns RM, Armstrong DV, Ax RL. Reproductive performance of Holstein cows receiving somatotropin. J Dairy Sci, 2000, 83(7): 1451-5
70. Manam S, Ledwith BJ, Barnum AB, Troilo PJ, Pauley CJ, Harper LB, Griffiths TG, 2nd, Niu Z, Denisova L, Follmer TT and others. Plasmid DNA vaccines: tissue distribution and effects of DNA sequence, adjuvants and delivery method on integration into host DNA. Intervirology, 2000, 43(4-6):273-81
71. McIntyre HD, Marechal DJ, Deby GP, Mathieu AG, Hezee-Hagelstein MT, Franchimont PP. Immunoreactive somatostatin in the rat ovary. Acta Endocrinol (Copenh), 1992, 126(6):553-8
72. Mor G, Yamshchikov G, Sedegah M, Takeno M, Wang R, Houghten RA, Hoffman S, Klinman DM. Induction of neonatal tolerance by plasmid DNA vaccination of mice. J Clin Invest, 1996, 98(12):2700-5
73. Mori T, Saito H, Ohno Y, Irahara M, Hosoi E, Saito S. Evidence for existence of somatostatin-like immunoreactivity with molecular heterogeneity in porcine ovaries. Acta Endocrinol (Copenh), 1984, 106(2):254-9
74. Naar EM, Bartke A, Majumdar SS, Buonomo FC, Yun JS, WagnerTE. Fertility of transgenic female mice expressing bovine growth hormoneor human growth hormone variant genes. Biol Reprod, 1991, 45:178-187
75. Nishiyama K, Sugimoto T, Kaji H, Kanatani M, Kobayashi T, Chihara K. Stimulatory effect of growth hormone on bone resorption and osteoclast differentiation. Endocrinology, 1996, 137(1):35-41
76. Owens PC, Conlon MA, Campbell RG, Johnson RJ, King R, Ballard FJ. Developmental changes in growth hormone, insulin-like growth factors (IGF-I and IGF-II) and IGF-binding proteins in plasma of young growing pigs. J Endocrinol, 1991, 128(3):439-47
77. Parker SE, Borellini F, Wenk ML, Hobart P, Hoffman SL, Hedstrom R, Le T, Norman JA. Plasmid DNA malaria vaccine: tissue distribution and safety studies in mice and rabbits. Hum Gene Ther, 1999, 10(5):741-58
78. Piedra PA. Clinical experience with respiratory syncytial virus vaccines. Pediatr Infect Dis J, 2003, 22(2 Suppl):S94-9
79. Pilling AM, Harman RM, Jones SA, McCormack NA, Lavender D, Haworth R. The assessment of local tolerance, acute toxicity, and DNA biodistribution following particle-mediated delivery of a DNA vaccine to minipigs. Toxicol Pathol, 2002, 30(3):298-305
80. Pillon D, Caraty A, Fabre-Nvs C, Lomet D, Cateau M, Bruneau G. Regulation by estradiol of hypothalamic somatostatin gene expression: possible involvement of somatostatin in the control of luteinizing hormone secretion in the ewe.Biol Reprod, 2004, 71(1):38-44
81. Pisetsky DS. Immune response to DNA in systemic lupus erythematosus. Isr Med Assoc J, 2001, 3(11):850-3
82. Pollak MN, Schally AV. Mechanisms of antineoplastic action of somatostatin analogs. Proc Soc Exp Biol Med, 1998, 217(2): 143-52
83. Rajkumar K, Kerr DE, Kirkwood RN, Laarveld B. Inhibitory action of somatostatin-14 on hormone-stimulated cyclic adenosine monophosphate induction in porcine granulosa and luteal cells. J Endocrinol, 1992, 134(2):297-306
84. Schalk JA, Mooi FR, Berbers GA, van Aerts LA, Ovelgonne H, Kimman TG. Preclinical and clinical safety studies on DNA vaccines. Hum Vaccin, 2006, 2(2):45-53
85. Schneider G, Fechner U. Advances in the prediction of protein targeting signals. Proteomics, 2004, 4(6): 1571-80
86. Shata MT, Stevceva L, Agwale S, Lewis GK, Hone DM. Recent advances with recombinant .bacterial vaccine vectors.Mol Med Today, 2000, 6(2):66-71
87. Sheets RL, Stein J, Manetz TS, Andrews C, Bailer R, Rathmann J, Gomez PL. Toxicological safety evaluation of DNA plasmid vaccines against HIV-1, Ebola, Severe Acute Respiratory Syndrome, or West Nile virus is similar despite differing plasmid backbones or gene-inserts. Toxicol Sci, 2006, 91(2):620-30
88. Shiau AL, Chu CY, Su WC, Wu CL. Vaccination with the glycoprotein D gene of pseudorabies virus delivered by nonpathogenic Escherichia coli elicits protective immune responses. Vaccine, 2001, 19(23-24):3277-84
89. Spencer GS, Hallett KG, Seddon A, Morris BA. Immuno-neutralization of maternal plasma oestradiol stimulates fetal growth in rats. Reprod Nutr Dev, 1986, 26(6): 1289-93
90. Spreng S, Viret JF. Plasmid maintenance systems suitable for GMO-based bacterial vaccines. Vaccine, 2005, 23(17-18):2060-5
91. Thomas AD, Murray JD, Famula TR, Oberbauer AM. Growth hormone and fertility in oMt1a-oGH transgenic mice. Reproduction, 2001, 122(4):537-44
92. Tran ND, Porada CD, Almeida-Porada G, Glimp HA, Anderson WF, Zanjani ED. Induction of stable prenatal tolerance to beta-galactosidase by in utero gene transfer into preimmune sheep fetuses. Blood, 2001, 97(11):3417-23
93. Tsang CH, Mirakhur KK, Babiuk LA, Griebel PJ. Oral DNA immunization in the second trimester fetal lamb and secondary immune responses in the neonate. Vaccine, 2007, 25(50):8469-79
94. Tseng J C, Levin B, Hurtado A, Yee H, Perez de Castro I,Jimenez M,Shamamian P,JinR,Novick RP,Pellicer A Meruelo D . Systemic tumor targeting and killing by sindbis viral vectors. NatBiotechnol, 2004, 22:70-77
95. Vernejoul F, Faure P, Benali N, Calise D, Tiraby G, Pradayrol L, Susini C, Buscail L. Antitumor effect of in vivo somatostatin receptor subtype 2 gene transfer in primary and metastatic pancreatic cancer models. Cancer Res, 2002, 62(21):6124-31
96. Welle S, Bhatt K, Pinkert CA, Tawil R, Thornton CA. Muscle growth after postdevelopmental myostatin gene knockout. Am J Physiol Endocrinol Metab, 2007, 292(4):E985-91
97. Wolf JJ, Wang L, Wang F. Application of PCR technology in vaccine product development. Expert Rev Vaccines, 2007,6(4):547-58
98. Wraith DC, Goldman M, Lambert PH. Vaccination and autoimmune disease: what is the evidence? Lancet, 2003, 362(9396): 1659-66
99. Xing Y, Wensheng L, Haoran L. Polygenic expression of somatostatin in orange-spotted grouper (Epinephelus coioides): molecular cloning and distribution of the mRNAs encoding three somatostatin precursors. Mol Cell Endocrinol, 2005, 241(1-2):62-72
100.Yoon HA, Aleyas AG, George JA, Park SO, Han YW, Lee JH, Cho JG, Eo SK. Cytokine GM-CSF genetic adjuvant facilitates prophylactic DNA vaccine against pseudorabies virus through enhanced immune responses. Microbiol Immunol, 2006, 50(2):83-92
101.Zaczek D, Hammond J, Suen L, Wandji S, Service D, Bartke A, Chandrashekar V, Coschigano K, Kopchick J. Impact of growth hormone resistance on female reproductive function: new insights from growth hormone receptor knockout mice. Biol Reprod, 2002, 67(4): 1115-24
102.Z.Roth, A.Arav, R.Braw-Tal, A.Bor,D.Wolfenson. Effect of Treament with Follicle-Stimulating Hormone or Bovine Somatotropin on the Quality of Oocytes Aspirated in the Autumn from Previously Heat-Stressed Cows. J Dairy Sci, 2002, 85:1398-1405