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1.
抑制素基因免疫对小鼠生殖的影响   总被引：32，自引：5，他引：27  
用编码抑制素α（1－32）的基因与pcDNA3.1真核表达质粒进行重组，构建抑制素基因疫苗NH。将30只2周龄ICR小鼠随机分为3组（每组10只），试验Ⅰ，Ⅱ组分别经脂质体介导肌肉注射iINH15μg()0μL,25μg(100μL)，对照组注射免疫空载体pcDNA3.115μg(100μL)。首次免疫后间隔3周加强免疫1次。小经pINH基因免疫后，用ELISA可从血浆中检测到抑制素抗体，表明pINH可在活体肌细胞内表达，表达产物具有制素抗原性，能激活免疫系统产生抑制素抗体。抑制素抗体P／N＞2视为阳性，阳性率为30％（6／20）。抑制素抗体阳性小鼠与阴性小鼠的产仔数，初生重和窝重等没有显著差异（P＞0．05）。抗体阳性组首次免疫后，雌鼠血浆FSH浓度略有升高，经过加强免疫后显著升高（P＜0．05）。由此可以看出，抑制素基因免疫可以诱导产生抗抑制素抗体，并促进促卵泡素的分泌。  相似文献   

2.
90只大鼠随机分为5组(n=18),分别肌肉注射10、50、100μg抑制素与乙肝表面抗原融合基因表达质粒(pCIS)、50μg空载体(pcDNA3.1)和100μL生理盐水。20 d后加强免疫1次,对每组中的12只大鼠进行2次加强免疫。结果发现抑制素抗体P/N值随着免疫次数的增加而提高。100μg剂量组2次和3次免疫的成熟卵泡发育数分别比对照组多6.9和7.5个(P<0.05)。3次免疫后大鼠成熟卵泡发育数比2次免疫后显著提高(35.2±2.73 vs 31.0±0.92,P<0.05)。抑制素基因免疫组的胎盘数和窝产仔数高于对照组(P>0.05),抗体阳性鼠高于阴性鼠(P<0.05)。pCIS 3次免疫后抗体阳性鼠的抗体水平与成熟卵泡发育数的相关系数为0.45(P>0.05),与胎盘数的相关系数为0.77(P<0.05)。pCIS免疫大鼠动情期和产后血浆FSH水平高于对照组,抗体阳性鼠的血浆FSH水平高于阴性鼠,其中2次免疫后阳性组动情期FSH浓度极显著高于阴性组(P<0.01)。这些结果表明,抑制素基因免疫大鼠可促进大鼠的卵泡发育,提高血浆FSH水平,为抑制素基因免疫大动物提供了试验依据。  相似文献   

3.
抑制素基因免疫大鼠的免疫应答与基因表达   总被引：1，自引：1，他引：1  
为了研究抑制素基因免疫对大鼠免疫应答和发情的影响及免疫后抑制素的组织分布,将60只大鼠分为5组,每只分别肌肉注射10（T1）、50（T2）、100μg pCIS（T3）,50μg pcDNA3.1（V）和生理盐水（S）。ELISA检测抑制素抗体水平,阴道涂片法检测大鼠的发情状况,免疫组化分析抑制素的组织分布。结果显示,不同剂量抑制素基因2次免疫10 d后抗体P/N值均显著升高（P〈0.05）,3次免疫10 d后T2和T3组抗体P/N值进一步显著升高（P〈0.05）,T3组抗体水平有高于T1和T2组的趋势（P〉0.05）;抑制素基因免疫对大鼠的发情无显著影响;3次免疫2周后心脏、肝脏、接种肌肉部位均未检出抑制素;卵巢、肾脏和垂体部位均检出抑制素;而脾脏部位,抑制素质粒免疫组检出抑制素,对照组则未检出抑制素。这些结果表明,免疫剂量和次数的增加没有导致大鼠产生明显的抑制素免疫耐受,抑制素基因免疫大鼠是相对安全的,抑制素的组织分布结果亦为抑制素基因免疫的作用机理研究提供了理论依据。  相似文献   

4.
将50只大鼠随机分为5组,分别注射0.2（T1）、0.6（T2）、1g/L（T3）pEGISI,100μg空载体pEGFP-N1（C1）和100μL生理盐水（C2）,以探讨在没有使用免疫佐剂的情况下,抑制素与GFP融合基因疫苗pEGISI免疫对大鼠卵巢和生殖激素的影响。研究结果,加强免疫能提高抗体P/N值,T3组在加强免疫期与对照组差异显著（P〈0.05）;加强免疫第2周时T3组抗体阳性鼠比例达40%,与T1和T2组差异均显著（P〈0.05）。T3组卵巢大小和卵泡数均显著高于对照组（P〈0.05）,但抗体阳性鼠大卵泡数与阴性鼠差异不显著（P〉0.05）。各剂量组促卵泡素和雌二醇均高于对照组,且在加强免疫第2周时T3组与对照组差异显著（P〈0.05）;各剂量组孕酮含量与对照组差异均不显著（P〉0.05）。结果表明,在没有使用免疫佐剂的前提下,抑制素pEGISI基因免疫可产生抗体,促进卵巢发育和FSH分泌。本试验条件下100μg是最佳免疫剂量。  相似文献   

5.
为提高动物繁殖力，分别用不同纯度的抑制素真核融合表达质粒pCISI，配以不同免疫佐剂后导入72头母黄牛体内。结果表明：①两次免疫后，产生抗抑制素抗体的黄牛占59．72％（4 3／72）；②pCISI能促进双大卵泡发育（68．75％，44／64），诱发排双卵（50．00％，32／64）；③加入佐剂的疫苗较未加佐剂的疫苗免疫效果明显，普鲁卡因佐剂显著优于脂质体（P〈0．05）；④纯度疫苗对牛的免疫效果略高于低纯度疫苗，但差异不明显（P〉0．05）。  相似文献   

6.
双拷贝抑制素基因疫苗pcISI的构建和表达及免疫   总被引：4，自引：2，他引：2  
为构建高免疫原性的卵泡抑制素（Inhibin,INH）DNA疫苗,将INH基因片段α1-32插入到pcIS中乙肝表面抗原（HBsAg）S基因第112-113氨基酸残基密码子之间,构建含2拷贝INH的融合表达质粒pcISI。酶切和测序鉴定表明,重组质粒pcISI构建成功。脂质体包裹法将pcISI转染HeLa细胞,ELISA检测表达产物的INH免疫反应原性。结果表明,融合目的基因在HeLa细胞获得表达,ISI融合蛋白具有比IS融合蛋白更强的INH抗原抗体反应性。将pcISI免疫6只大鼠后,5/6的大鼠产生了抗INH抗体,抗体P/N值在免疫后第2-6周高于pcIS免疫组。这些结果表明,所构建的质粒pcISI可以表达抑制素,表达产物具有较强的免疫原性。  相似文献   

7.
将表达完整猪生殖与呼吸综合征病毒（PRRSV）GP5和融合表达泛素与GP5成熟肽的真核重组表达质粒（pCI—GP5和pcDNA—U—GP5）分别肌肉注射免疫BALB／c小鼠，共免疫3次。采用间接ELISA法检测血清中抗GP5的抗体水平，MTT法检测脾淋巴细胞的增殖能力，LDH释放试验检测脾NK细胞的杀伤活性，并通过流式细胞术检测脾淋巴细胞亚群的比例。结果发现，2种DNA重组质粒免疫后均可诱导小鼠产生体液免疫和细胞免疫。三免后第5周，pCI—GP5质粒免疫组诱导产生的抗GP5抗体水平及NK活性明显高于pcDNA—U—GP5质粒免疫组；而pcDNA—U—GP5免疫组的淋巴细胞增殖指数及CD4%-CD8^＋T细胞比例明显高于pCI—GP5组。表明，GP5基因可诱导产生较高的免疫反应，泛素与GP5的融合表达可增强PRRSVGP5诱导的细胞免疫反应。  相似文献   

8.
将90只大鼠随机分为5组,分别注射10μg.100μL-1 pcISI(T1)、50μg.100μL-1 pcISI(T2)、100μg.100μL-1(T3)pcISI、100μg空载体pcMV-S(V)和100μL生理盐水(S),以探讨在不使用免疫佐剂的情况下,不同剂量的双拷贝抑制素pcISI基因免疫对大鼠卵泡发育、产仔和生殖激素的影响。结果表明,加强免疫能提高抗体P/N值,加强免疫后各剂量组平均抗体P/N值大于2,T3组抗体P/N值显著高于T1和T2组(P<0.05)。T3组成熟卵泡数显著高于对照组(P<0.05),且抗体阳性鼠成熟卵泡数比阴性鼠多12.45个(P<0.05)。除了T1组外,其他2个剂量组产仔数和胎盘数均高于对照组(P<0.05)。抗体阳性鼠胎盘数比阴性鼠多5.09个(P<0.05),产仔数比阴性鼠多5.39个(P<0.05)。抑制素pcISI基因免疫大鼠后促卵泡素(FSH)、雌二醇(E2)和孕酮(P4)含量均高于对照组,T3组的FSH含量与对照组差异显著(P<0.05),T3组的E2含量与对照组差异极显著(P<0.01),各剂量组P4含量与对照组差异均不显著(P>0.05)。这些结果说明,在没有免疫佐剂的前提下,双拷贝抑制素pcISI基因免疫可产生抗体,促进FSH分泌和卵泡发育,增加产仔数。本试验条件下100μg.100μL-1是最佳免疫剂量。  相似文献   

9.
选择40头南阳黄牛，用3．0毫克抑制素基因重组质粒（PCIS，免疫组，n=30）或同剂量的生理盐水（对照组，n=10）间隔21天注射2次，分别在首次免疫当天、首次免疫后第10天和第21天、加强免疫后第10天和第45天收集血清，应用放射免疫法（RIA）测定处理后不同时期血清中激素水平。结果表明：在5个采血时间点内．加强免疫后第10天抗体阳性牛比率最高．为37．5％，与其他采血时间点有显著差异。免疫组促卵泡素（FSH）平均含量比对照组高，但差异不显著；17-β雌二醇（E2）和孕酮（P4）水平在加强免疫后2组有显著性差异。  相似文献   

10.
选择32只60日龄雌性性成熟大鼠分为4组，分别肌肉注射pcDNA-DPPISS-DINH（试验组1）50μg，poD—NA—DPPISS-DINH—mC3d3（试验组2）50μg，空载体pcDNA3．1（对照组1）50μg，生理盐水（对照组2）1mL。首次免疫20、40d后各组分别同剂量同样免疫程序加强免疫注射1次。结果发现，试验1和2组卵巢的重量都显著高于对照组1和2，但试验1和2组之间的差异不显著（P〉0．05）；试验2组卵巢成熟卵泡发育个数和FSH的浓度显著高于试验1组和对照1和2组（P〈0．05）；加强免疫第40天，试验1、2组的E1的浓度分别显著高于本试验组免疫前第0天，免疫后第20天的激素浓度；试验组与对照组在各个免疫试验时间LH的浓度在统计上差并不显著（P〉0．05）。结果证明，应用重组抑制素真核表达质粒pcDNA-DPPISS-DINH—mC3d3（试验组2）免疫动物可以促进大鼠卵泡的发育，可提高血浆FSH、E2水平，为抑制素基因免疫大动物提供了试验依据。  相似文献   

11.
1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.  相似文献   

12.
Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.  相似文献   

13.
The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.  相似文献   

14.
The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.  相似文献   

15.
Comparison of different tests to diagnose feline infectious peritonitis   总被引：1，自引：0，他引：1  
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16.
An experiment was conducted to study disappearance of C14 to C18 fatty acids, lag times and biohydrogenation (BH) rates of C18 fatty acids of ground soya bean and canola seeds in situ. Three ruminally fistulated Dallagh sheep were used to determine ruminal BH of unsaturated fatty acids (UFAs). Differences in the disappearance of fatty acids through the bags and lag times were observed between the oilseeds. We saw that the longer the incubation time of the oilseeds in the rumen, the lower the content of C18:2 and C18:3. Significantly higher lag times for both C18:2 and C18:3 were observed in ground canola compared to ground soya bean. BH rates of C18:2 and C18:3 fatty acids in soya bean were three times higher than those of canola. These results suggest that the fatty acid profile of fat source can affect the BH of UFAs by rumen micro‐organisms. So that UFAs of canola had higher ability to escape from ruminal BH. It seems that fatty acid profile of ruminant products is more affected by canola seed compared to soya bean seed.  相似文献   

17.
Serum biochemical values and the wet weight to dry weight ratios of tissues were determined in yaks in Shandan county of Gansu province. The liver, kidney, heart and muscle contents of seven elements in yaks were also analysed. Most serum biochemical values were similar to those of cattle, camels and sheep, but the calcium concentration was considerably above the normal range for other ruminants. The liver contained the highest concentrations of copper, manganese and cobalt and the kidney of selenium, iron and calcium.Abbreviations Alb albumin - ALT alanine aminotransferase - AST aspartate aminotransferase - BUN blood urea nitrogen - Chol cholesterol - Glob globulin - -GT -glutamyl transpeptidase - IP inorganic phosphorus - TP total protein  相似文献   

18.
A knowledge of the microbiological status of milk and of the different structures in the mammary glands has great importance in elucidating the pathogenesis of mammary gland infections. The objective of this study was to evaluate the microbiological status of various structures in the mammary glands from naturally infected dairy cows following slaughter. A total of 94 samples of milk, 184 samples of mammary parenchyma, 168 samples of gland cisterns, and 168 samples of teat cisterns were collected for microbiological examination. Microorganisms were detected in 59.9% of all samples, 67.0% of the milk samples, 70.1% of the mammary parenchymas, 55.9% of the gland cisterns and 48.8% of the teat cistern samples. When all samples were considered, coagulase-negative Staphylococcus were the most prevalent (35.7%) followed by coagulase-positive Staphylococcus (12.2%), Corynebacterium bovis (2.4%), Prototheca sp. (1.9%), and Streptococcus dysgalactiae (1.5%). There was a significantly higher occurrence of microorganisms in the milk and mammary parenchyma compared to the gland cisterns and teat cisterns. This revised version was published online in July 2006 with corrections to the Cover Date.  相似文献   

19.
Four grass plots were sequentially contaminated with goat faeces containing known numbers of unembryonated eggs of predominantly Haemonchus contortus and Trichostrongylus spp. between October 1982 and April 1983. Four other plots were similarly contaminated with sheep faeces between February and May 1987. An additional plot was repeatedly contaminated with sheep faeces from February to April 1987. Populations of free-living stages in faeces and of infective larvae (L3) in the herbage were subsequently monitored until the end of April and June of 1983 and 1987 respectively. During February and May 1987 two control cultures of sheep faeces were incubated in the laboratory at 25°C–30°C and at a constant temperature of 50°C and the free-living development was also monitored. L3 developed very readily in the faeces cultured at 25°C–30°C and in those spread on a grass plot in October, at the end of the wet season, but developed less on the plot contaminated in May at the start of the wet season. Worm eggs in faeces deposited on plots during the hot dry season (December to April) or incubated at 50°C died and disintegrated after 24–48 h exposure to the high environmental temperatures. The results indicate that it is unlikely that gastrointestinal nematodes of sheep and goats can develop or survive on open pasture during the dry season in the Nigerian derived savanna zone.  相似文献   

20.
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