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本试验的目的是评估抗坏血酸对Ross—08鸡体外精子质量的影响．采集8只青年期Ross-308肉种公鸡的精液将将它们混合在一起。混合后的精液用改良林格氏液稀释并添加5个处理水平的抗坏血酸（w／v）（0、0．5％、1％、2％和3％）。4℃贮存，并在贮存后的第0、3、6、10和24h对处理组进行精子活力、活率和畸形率的测定。试验结果表明，在贮存后的第0、3和6h，添加抗坏血酸的处理组与对照组精子质量参数无显著差异。在贮存后的第10h．添加1％抗坏血酸处理组精子活力显著高于对照组和其他处理组，添加了0、0．5％、2％和3％抗坏血酸处理组间没有显著差异：在贮存后的第24h，1％抗坏血酸处理组的精子活力与添加1％和2％抗坏血酸处理组精子活率均高于对照和其他处理组：1％抗坏血酸处理组精子畸形率显著低于对照组和其他处理组：在贮存后的第24h，添加3％的抗坏血酸处理组精子畸形率显著高于对照和其他处理组。 相似文献
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应用细胞骨架抑制剂细胞松弛素B和秋水仙素处理兔成熟卵母细胞，然后采用电激活的方法激活兔卵母细胞，观察细胞骨架抑制剂对兔卵母细胞孤雌激活和孤雌发育的影响，结果表明：应用不同的电激活液，即甘露醇、Zimmerman氏和山梨醇对兔卵母细胞的孤雌激活效果无显著性差异（P〉0．05）；用7．5μg·mL^-1细胞松弛素B处理卵母细胞后孤雌激活，其2-细胞胚率（82．2％）和囊胚发育率（43．4％）与对照组（76．4％和51．5％）相比无显著性差异（P〉0．05）；用1μg·mL^-1秋水仙素处理兔卵母细胞，兔卵母细胞孤雌激活后的2-细胞胚率（64．9％）和囊胚发育率（23．8％）明显下降，与对照组相比存在显著性差异（P〈0．05）；用细胞松弛素B和秋水仙素共同处理的2-细胞胚率（62．5％）和囊胚发育率（30．9％）与秋水仙素单独处理的结果无显著性差异（P〉0．05）。因此，秋水仙素对兔卵母细胞的孤雌激活影响比CB更大。通过免疫荧光和激光扫描共聚焦显微镜观察发现，细胞骨架抑制剂处理后，经孤雌激活获得的囊胚中，微丝和微管结构未见异常。 相似文献
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活蛹缫丝能直接选拔茧丝质优良的个体继代,是家蚕品种改良和选育中提高丝质的有效方法。试验表明：用不同浓度的硅酸钠＋0．1％氢氧化钠的混合液作浸茧液进行活蛹缫丝时,对家蚕的蛹体健康性和产卵量有不同程度的影响。0．4％～0．6％硅酸钠＋0．1％氢氧化钠处理区健蛹率保持在96．67％～98．33％,与对照差异不显著;产卵量比对照减少13．54％～16．44％,对家蚕蛹体影响较小,宜选作活蛹缫丝浸茧液。浓度0．7％以上的硅酸钠＋0．1％氢氧化钠的混合液作浸茧液,对家蚕蛹体健康性及产卵量均有显著影响。 相似文献
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同期发情处理方法对受体牛胚胎移植效果影响的研究 总被引：4，自引：0，他引：4
试验设计3个组．分别采用CIDR＋PGF2α法、PGF2α一次注射法和PGF2α二次注射法3种不同的处理方法对120头西门塔尔杂交受体牛进行同期发情处理。结果表明：CIDR＋PGF2α法同期发情处理的受体牛黄体不合格率达47．4％．显著高于其他两组（P〈0．05）;胚胎移植产犊率为45．0％,分别比PGF2α一次注射法和PGF2α二次注射法低10．2和13．1个百分点。PGF2α二次注射法同期发情处理后的受体牛黄体不合格率仅20．5％,低于PGF2α一次注射法（P〉0．05）,显著低于CIDR＋PGF2α法（P〈0．05）;胚胎移植产犊率达58．1％,均高于其他两组。本试验初步显示：采用PGF2α二次注射法进行同期发情处理受体牛效果好,成本较低,操作方便,产犊率也较高。 相似文献
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选1日龄商品代艾维因混合健雏480只，按单因子随机试验设计分为4组，每组4个重复，每重复30只进行试验。对照组饲喂基础日粮 5mg／kg维基尼亚霉素；试验l，2，3组鸡21日龄前分别饲喂基础日粮 0．1％生命素 0．5％迈克活菌酶；基础日粮 500玎骧／kg糖萜素 0．5％迈克活菌酶；基础日粮 300mg／／kg糖萜素 0．3％迈克活菌酶 0．06％生命素。21日龄后分别饲喂基础日粮 0．1％生命素 0．3％迈克活菌酶；基础日粮 300叫kg糖萜素 0．3％迈克活菌酶；基础日粮 200mg／kg糖萜素 0．2％迈克活菌酶 0．05％生命素。各组基础日粮完全相同。试验结果表明：3个试验组供试鸡的体增重分别比对照组提高10．24、9．61和12．48％．料重比分别降低7．50、7．08和11．67％，每只鸡实际赢利分别增加0．5ll、0．671和1．019元，生物学综合评定值分别为106．12、105．75和108．56。不同无公害添加剂组合对维基尼亚霉素具有显著替代作用。 相似文献
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去甲肾上腺素和睾酮对低温保存犬精子的影响 总被引：2，自引：0，他引：2
本试验就去甲肾上腺素和睾酮对低温保存犬精子的影响液进行了研究。试验结果表明：①稀释液—Tris-葡萄糖稀释液（Tris3．04％,柠檬酸1．7％,葡萄糖1．25％,卵黄20％）中加入去甲肾上腺素后,犬精液低温保存的效果显著优于对照稀释液和加入睾酮的稀释液,在4℃保存4d后,加入去甲肾上腺素的稀释液中的精子活率高达0．6500-0．01378,精子活率〉0-3的存活时间达8．3333±2．5819d,而对照稀释液和加入睾酮的稀释液中的精子活率分别只有0．45±0．16和0．36±0．11,精子活率〉0．3的存活时间分别为5．67±1．75和4．67±1．03。②对去甲肾上腺素的不同的4种添加剂量研究后发现,100μL稀释液中加入15μL去甲肾上腺素时犬精子低温保存的效果最好。 相似文献
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试验旨在探究不同pH的弱酸性环境常温稀释液对于猪精液常温保存的影响。通过测定不同pH（PH为6．2、6．3、6．4、6．5、6．6、6．7）的稀释液条件下猪精子的活率、质膜完整率和顶体完整性来检测对猪精液的保存效果。结果表明,稀释48h后,pH为6．4和6．5的稀释液中精子活率、质膜完整性和顶体完整性都分别出现降低,明显低于对照组（P〈0．05）。pH为6．2时,稀释液中精子的活率、质膜完整性和顶体完整性显著降低（P〈0．01）,不同PH的稀释液稀释后精液的品质在24h后开始出现明显的下降（P〈0．05）。试验表明,适宜猪精液常温保存的稀释液的弱酸性环境PH为6．4和6．5,在24h内保存效果较好。 相似文献
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小鼠睾丸支持细胞分离培养及生物学特性鉴定 总被引：1，自引：0，他引：1
为了揭示睾丸支持细胞在精子发生及睾丸免疫豁免中的生物学作用,采用胰蛋白酶、胶原酶分步消化法和1/3浓度的Hank′s低渗液、50mmol/L Tris-HC(l pH7.1)液分别低渗处理及油红O脂质染色法对16～22d的幼鼠睾丸进行分离、纯化和生物学特性鉴定。结果表明:分离、纯化后的睾丸支持细胞损伤少、活率高、睾丸支持细胞占培养细胞总数的90%以上;根据形态学观察及油红O脂质染色法鉴定,睾丸支持细胞的生长状态良好,细胞突起很多,核仁清晰,胞质中可见大小不等的红色空泡状脂质小滴。 相似文献
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小鼠睾丸支持细胞体外分离培养的研究 总被引：1，自引：0，他引：1
为了研究一种能快速、高效地分离、纯化小鼠睾丸支持细胞的方法,试验采用颈部脱臼的方法处死3周龄的小白鼠,取其睾丸并去除附属组织(血管、白膜脂肪等)后剪碎,用Ⅳ型胶原酶和胰蛋白酶分步消化制备细胞悬液进行培养,台盼蓝染色以鉴定细胞的活率;再对细胞悬液进行低渗溶液处理并利用支持细胞贴壁而其他细胞不贴壁的特性对其进行分离、纯化;最后对支持细胞采用H.E.和油红O染色的方法进行鉴定,观察其形态、结构和生长增殖情况。结果表明:该方法能够有效地分离、纯化以及培养小白鼠睾丸支持细胞。 相似文献
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为研究褪黑素对雏鸡睾丸支持细胞增殖的影响,本实验以原代分离培养并鉴定的雏鸡睾丸支持细胞为研究对象,经不同浓度(1、10、100、1000 nmol/L)褪黑素处理细胞48 h后,使用Cell Counting Kit-8(CCK-8)、EdU和实时荧光定量PCR(RT-qPCR)检测雏鸡睾丸支持细胞的细胞活力、细胞增殖能力和增殖相关基因的表达。结果表明:与对照组相比,添加1000 nmol/L褪黑素可提高细胞活力和细胞增殖能力(P<0.05),上调PCNA、Cyclin B和Cyclin D基因的表达(P<0.05),下调P21基因的表达(P<0.05)。结果显示褪黑素可以促进支持细胞增殖。 相似文献
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The influence of selected techniques of bovine leukocyte isolation on their viability and metabolism
Urban-Chmiel R Lisiecka U Chłopaś A Kurek Ł Wernicki A 《Polish journal of veterinary sciences》2011,14(4):663-665
The aim of the study was to assess the effect of selected isolation methods on the viability and metabolism of bovine leukocytes. The cells were isolated using a Ficoll 1077, Histopaque 1083 gradient and osmotic shock method, and Ficoll or Histopaque with osmotic shock. Evaluation were made of the total number of cells, viability after isolation and in 24h culture on RPMI 1640 medium and metabolism with NBT reduction assay. Microscopic and cytometric evaluation of the leukocytes revealed that the isolation methods applied had an influence on their number and viability. Based on the results it can be concluded that isolation methods of cells in a Histopaque or Ficoll yield highly pure cell fractions with high viability. 相似文献
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Mast cell tumor destruction by deionized water 总被引：2，自引：0，他引：2
R L Grier G Di Guardo C B Schaffer B Pedrosa R Myers D F Merkley M Thouvenelle 《American journal of veterinary research》1990,51(7):1116-1120
In a controlled study, malignant murine P815 mastocytoma cells exposed in vitro to distilled and deionized water died as a result of progressive swelling, degranulation, and membrane rupture. A 90% mean cell death occurred when cells obtained directly from culture were exposed to deionized water for 2 minutes. Of 6 cryopreserved malignant murine cell lines, which included Cloudman S91 melanoma, CMT-93 rectum carcinoma, MMT-06052 mammary carcinoma, and S-180 Sarcoma, only P815 mastocytoma and YAC-1 lymphoma were significantly (P less than 0.05) affected by hypotonic shock; Cloudman S91 melanoma cells were the most resistant. Mastocytoma cells were selectively killed by hypotonic solution, and lymphoma cells were also killed by isotonic saline solution. Local mast cell tumor (MCT) recurrence and percentage survival were evaluated in 12 cats (21 MCT) and 54 dogs (85 MCT) subjected to surgery alone or local infiltration of deionized water as an adjunct to surgery. Of all 16 incompletely excised MCT in cats, there was no local recurrence following injection. Four mast cell tumors (2 cats) regressed after being injected in situ. In dogs with clinical stage-I MCT, local recurrence was detected in 50% (5/10), but with injection after incomplete excision, local MCT recurrence was significantly (P less than 0.05) less (6.6%, 1/15). Percentage recurrence was significantly (P less than 0.05) less and survival significantly greater when incompletely excised grade-II MCT were injected. Mean follow-up period after surgery in cats and dogs was 35 and 23.4 months, respectively. 相似文献
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为研究一种简单快速分离蒙古马睾丸支持细胞并保证其活性的基本方法,本试验采集2岁蒙古马睾丸组织于低温环境下进行机械分离,将1~3 g睾丸组织剪碎,采用重力沉淀法去除游离的红细胞和间质细胞;使用0.1%胶原酶Ⅳ和0.25%胰蛋白酶+EDTA逐步进行组织消化,并用含有10%胎牛血清的培养基进行细胞培养;在接种后采用差异贴壁法分离纯化支持细胞,使用Tris-HCl低渗法去除杂质细胞,并采用碱性磷酸酶（AKP）染色、HE染色、实时荧光定量PCR方法进行鉴定。结果显示,支持细胞贴壁性能较强,培养24 h后大多数细胞开始贴壁,细胞形状呈椭圆形;培养48 h后胞质增多,折光性变强;培养3~4 d细胞胞质展开紧密连接,此时细胞呈三角形,有明显的核仁,培养6~7 d细胞生长状态进入稳定期。实时荧光定量PCR结果显示,GATA4和GDNF基因在培养细胞中极显著表达（P<0.01）,AKP染色支持细胞呈阴性表达,表明分离培养的细胞确为支持细胞。本试验使用机械分离法与两步酶消化法处理组织,可快速高效地获得睾丸支持细胞,成功构建了蒙古马睾丸支持细胞体外培养方法。 相似文献
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SONG Lianjie CUI Yingying ZHAO Yiping BAI Dongyi REN Xiujuan TE Rigele DUGARJAVIIN Manglai LI Bei 《中国畜牧兽医》2007,47(9):2751-2758
To study a simple and rapid separation of testis Sertoli cells in Mongolian horses and ensure their activity,the testis tissue of two-year-old Mongolian horses was mechanically isolated under a low temperature environment in this experiment,and 1-3 g of testis tissue was chopped,the free red blood cells and interstitial cells were removed by gravity precipitation method.0.1% collagenase Ⅳ and 0.25% trypsin-EDTA were used for tissue digestion, and cells were cultured in medium containing 10% fetal bovine serum.After inoculation,the purified Sertoli cells were isolated and purified by differential sticking method,and the impurity cells were removed by Tris-HCl infiltration method,and were identified by alkaline phosphatase (AKP) staining,HE staining and Real-time quantitative PCR.The results showed that most of the cells began to adhere to the wall after culture 24 h,and the shape of the cells was oval.After culture 48 h,the cytoplasm increased and the refraction became stronger.After culture 3-4 d,the cytoplasm of the cells expanded into tight junction.At this time,the cells were triangular with obvious nucleoli.After culture 6-7 d,the cells entered the stable phase.Real-time quantitative PCR results showed that the expression of GATA4 and GDNF genes were extremely significantly expressed in cultured cells (P<0.01).AKP staining supported the negative expression in Sertoli cells,indicating that the isolated cultured cells were indeed Sertoli cells.In this experiment,tissue was treated by mechanical separation and two-step enzyme digestion,and testicular support cells were obtained quickly and efficiently,the method of culturing Sertoli cells in Mongolian horses testis in vitro was successfully constructed. 相似文献
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Schneider N Lejeune JP Deby C Deby-Dupont GP Serteyn D 《Veterinary journal (London, England : 1997)》2004,168(2):167-173
Ischaemia and reperfusion are suspected to alter chondrocyte metabolism. Here, we studied the effects of three oxygen (O2) tensions on the viability of equine articular chondrocytes isolated from the cartilage of the distal interphalangeal joint of horses. Chondrocytes were cultured in alginate beads under 1%, 5% or 21% gas phase O2 concentration for 14 days, cellular growth kinetics were measured (n=6), and the cells were observed by light microscopy after staining for necrotic and apoptotic cell detection. For information about the metabolic status, the intracellular adenosine triphosphate (ATP) content was measured. The number of chondrocytes remained stable for the first eight days, then decreased especially at 1% and 21% O2. At 21% O2, normal cells decreased and necrotic cells increased at the end of the 14 day-period. No significant variations were found at 5% O2 except for a decrease in necrotic cells at day 14. Most apoptotic cells were found at 1% O2 from days 5 to 11, and normal cells decreased during the same period. But an unexpected increase in normal cells and decrease in apoptotic cells were observed at day 14. The intracellular ATP content remained stable. It was concluded that, in a three-dimensional culture model of equine articular chondrocytes, O2 tension affected the viability of the cells after an 11-day period, with the most important effects observed at 21% and 1% O2 conditions. 相似文献
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花生四烯酸对日本沼虾肝胰腺细胞脂质代谢基因表达的影响 总被引：1，自引：0，他引：1
本试验旨在评价细胞培养液中花生四烯酸(arachidonic acid,ARA)浓度对日本沼虾肝胰腺细胞活力及脂质代谢相关基因表达的影响。分离日本沼虾肝胰腺细胞,使用M199完全培养液培养5 d后换成含ARA的培养液,ARA浓度分别为0(ARA1)、50(ARA2)、100(ARA3)、200(ARA4)和1 000μmol/L(ARA5),测定12和24 h时脂质代谢相关基因的表达水平,以及24h时细胞活力。结果表明:原代肝胰腺细胞使用完全培养液时,生长状况良好,能存活15 d左右;ARA5组24 h时细胞活力显著低于ARA1和ARA2组(P0.05);高浓度的ARA降低了12和24 h时Δ4脱饱和酶(Δ4 FAD)、Δ6脱饱和酶(Δ6 FAD)、碳链延长酶6(Elovl6)、B类Ⅰ型清道夫受体(SR-BⅠ)、脂肪酸结合蛋白10(FABP10)、乙酰辅酶A结合蛋白(ACBP)基因表达水平;ARA作用12 h时,ARA2组SR-BⅠ基因表达水平显著高于其余各组(P0.05),ARA2和ARA3组FABP10基因表达水平显著高于ARA1和ARA5组(P0.05),ARA3组ACBP基因表达水平显著高于其余各组(P0.05);ARA作用24 h时,ARA2组SR-BⅠ、FABP10和ACBP基因表达水平显著高于其余各组(P0.05)。由此可见,细胞培养液中ARA浓度会影响日本沼虾肝胰腺细胞活力及脂质代谢相关基因的表达,过高的ARA浓度(1 000μmol/L)会降低细胞的活力,适宜的ARA浓度(50~100μmol/L)可促进脂肪酸脱饱和酶、碳链延长酶及脂肪酸转运相关基因的表达。 相似文献