牛卵母细胞的孤雌激活研究

本文探讨了透明质酸酶(HYA)的浓度和作用时间对牛体外成熟卵母细胞脱卵丘的影响,不同强度电脉冲 乙醇 6-DMAP对牛体外成熟卵母细胞的孤雌激活作用以及和乙醇 6-DMAP激活的比较。对电融合脉冲强度、脉冲次数进行了优化。结果表明:卵母细胞成熟培养22h,用0.20%HYA作用10min或0.50%HYA作用5min效果较好。在激活电压1.6kV/cm、20μs/次、间隔1s,脉冲次数为1次的条件下的激活效果较好,在此条件下与乙醇 6-DMAP激活相比较,孤雌激活胚胎的囊胚率分别为19.6%和26.9%,差异不显著。     

兔卵母细胞孤雌激活方法的研究

对兔卵母细胞的孤雌激活方法进行探讨,为兔的体细胞核移植奠定良好的技术平台。体外成熟培养20～22h的兔卵母细胞随机分组,分别进行以下试验：采用不同脉冲电压、脉冲次数和脉冲时间,电激活兔卵母细胞,摸索最佳电激活参数;比较不同的激活方法处理免卵母细胞的效果,Ⅰ：Ion5min＋6-DMAP3h,Ⅱ：Ion5min＋CHX3h,Ⅲ：Ion5min＋6-DMAP联合CHX3h,Ⅳ：电激1次联合6-DMAP＋CHX3h;优化6-DMAP与CHX作用时间。结果显示：当电激参数为：电压40V／mm,脉冲时间20μs和脉冲次数3次时,兔孤雌胚的分裂率和囊胚率最高,可达80．64％和14．51％;采用第Ⅳ种方法激活兔的卵母细胞,胚胎的分裂率和囊胚率（80．64％,13．71％）均显著高于Ⅰ组（66．67％,8．6％）和Ⅱ组（43．21％,1．24%,P〈0．05）,略高于第Ⅲ组（77．59％,12．07％）;当兔孤雌胚分别在含6-DMAP和CHX的CM中培养1。h和3h时,1h处理组在分裂率（80．68％vs77．59％）和囊胚率（15．91％vs12．07％）上都略高于3h处理组。结果表明：兔卵母细胞经Ion处理5min,或先电激1次（参数为电压40V／mm,脉冲时间20μs,脉冲3次）,再用2mmol／L6-DMAP＋5mg／LCHX处理1～3h,均可获得较好的激活效果,有利于兔孤雌胚的发育。     

不同激活方法对猪体外成熟卵母细胞孤雌发育的影响

通过系统探讨不同孤雌激活方法对猪卵母细胞体外激活后发育效果的影响,比较了乙醇（Ethanol,EH）、离子霉素（Ionomycin,Ion：5μmol/L）、氯化锶（Strontium chloride hexahydrate,Sr^2＋：10 mmol/L）、6-二甲氨基嘌呤（6-dimethylaminopurine,6-DMAP：2 mmol/L）和放线菌酮（cycloheximide,CHX：10 mg/L）对猪卵母细胞激活发育的效果。结果表明：（1）9%EH激活处理10 min效果好于15 min;（2）使用9%EH激活处理10 min,再结合CHX、6-DMAP、Sr^2＋、CHX＋Sr^2＋、Sr^2＋＋6-DMAP、CHX＋6-DMAP或CHX＋6-DMAP＋Sr^2＋组处理3～4 h,以EH＋6-DMAP组效果最好,分裂率及囊胚率分别达到82.86%和22.86%;（3）在使用化学激活（Ion＋6-DMAP组和9%乙醇10 min＋6-DMAP组）和电激活（50 V/mm,50μs,2t）的方法中,Ion法激活猪卵母细胞效果较好,囊胚率达到37.50%;（4）卵母细胞包被的卵丘细胞层数不同对卵母细胞成熟激活有显著的影响,卵丘细胞层数4～6层和多于6层的卵丘-卵母细胞复合体孤雌激活的分裂率和囊胚率分别为（68.99%,32.56%）和（75.36%,37.68%）,2组之间差异不显著（P〉0.05）;但其显著高于其他组（P〈0.05）,这2组细胞在猪孤雌激活发育研究中是最佳的实验研究材料。     

不同化学激活剂对小鼠卵母细胞的激活效果

为探讨不同激活剂的卵母细胞孤雌激活效果及激活胚体外发育情况,用乙醇、CaA23187、SrCl2、6-DMAP及CB分别对小鼠卵母细胞进行了激活处理.结果显示,70 mL/L乙醇刺激小鼠卵母细胞时,以激活5～7 min的激活效果及孤雌胚发育较好,桑囊胚发育率可达29.11%;用SrCl2激活小鼠卵母细胞时,6～10 mmol/L为最佳处理浓度,3～6 h为最佳激活时间,桑囊胚发育率可达16.67%;以70 mL/L乙醇激活5 min,再用2 mmol/L 6-DMAP＋5 μg/mL CB激活3 h效果最佳,桑囊胚发育率高达35.77%.研究证实,小鼠卵母细胞经乙醇、6-DMAP和CB等复合激活后能较好地发育.     

水牛卵母细胞孤雌激活及孤雌胚与体外受精胚发育比较

对MII期水牛卵母细胞进行人工诱导激活可以帮助人们间接判断体外成熟卵母细胞质量的优劣。并且，卵母细胞的充分激活也是提高核移植效率的关键因素之一。试验比较了水牛卵母细胞体外成熟时间对孤雌激活胚发育能力的影响以及不同化学激活方法对水牛卵母细胞孤雌激活效果的影响，并在相同条件下，对孤雌激活胚与体外受精胚的发育能力进行了比较。结果表明，水牛卵母细胞体外成熟27小时或30小时的囊胚发育率（19．0％或17．7％）明显高于体外成熟21小时或24小时的囊胚发育率（12．3％或13．8％）；Ion联合6-DMAP激活水牛卵母细胞的效果优于其他几组激活方法；在相同条件下，孤雌激活胚与体外受精胚的发育能力存在着差异，其中卵裂率差异不显著，但孤雌激活胚的囊胚发育率显著高于体外受精胚（13．0％vs21．7％）。     

小鼠卵细胞体外成熟培养及孤雌激活

[目的]利用小鼠卵母细胞体外成熟培养及3种激活方法（乙醇激活、氯化锶激活、乙醇-氯化锶激活）对小鼠卵母细胞进行了孤雌激活研究。[方法]小鼠经注射孕马血清促性腺激素（PMSG）48h后,摘取卵巢获得3级未成熟卵母细胞．在成熟培养液（M199100mL＋丙酮酸钠2．2mg＋抗100IU／mL＋LH2IU／mL＋FCS10％）中对小鼠未成熟卵细胞进行体外成熟培养,获得的成熟卵母细胞分别在乙醇、氯化锶、乙醇一氯化锶不同激活剂中激活,[结果]A级卵母细胞成熟率为79％．B级卵母细胞成熟率为70％,C级卵母细胞成熟率为55％。A级卵母细胞使用氯化锶激活率可达80．0％,为最佳方法。[结论]A级卵母细胞成熟培养效果最好。孤雌激活氯化锶激活效果高于乙醇．．     

不同方法对滩羊卵母细胞孤雌激活及重构胚发育影响的研究

旨在探索宁夏滩羊卵母细胞孤雌激活及胚胎培养条件,并建立转Cherry基因滩羊体细胞重构胚的融合与体外培养体系。试验分析了3种激活方法即电激活、Ionomycin结合6-DMAP与电激活结合6-DMAP对成熟的滩羊卵母细胞激活的影响,以及3种胚胎培养液mSOFaa、M16和KSOM的培养筛选,并优化了滩羊体细胞重构胚的融合与体外培养条件。结果表明:在卵母细胞孤雌激活中,最适电压为1 600V/cm,获得卵裂率和囊胚率分别为58.5%和19.5%;5μmol离子霉素结合2 mmol 6-DMAP能有效地激活成熟的滩羊卵母细胞,卵裂率为81.0%,囊胚率为26.2%;并发现mSOFaa胚胎培养液的卵裂率和囊胚显著优于M16和KSOM培养液;在转基因重构胚融合中发现在电压1 800V/cm、脉冲时间10μs、脉冲次数2次和间隔时间为1s的条件下,卵裂率和囊胚率为40.0%、21.4%。本试验建立的滩羊孤雌激活和转基因重构胚融合与体外培养体系为宁夏滩羊的分子育种奠定了基础。     

兔卵母细胞的孤雌激活

从电脉冲次数、脉冲强度、脉冲间隔时间三方面对兔卵母细胞的孤雌激活进行了研究。结果表明，注射HCG后16h的MⅡ期卵，用1．4kV／cm、60μs的直流脉冲电激，脉冲次数分为1、2、4、8次4组，每次间隔30min，随着电激活次数的增加，活化率从46．5％增加到95．8％(P＜0．01)，但是电激活8次后，桑椹胚率却从63．6％下降列21．2％，囊胚率从23．6％下降列2．8％(P＜0．01)；用60μs的直流脉冲电激3次，每次间隔30min，分为0．4、1．2、2．4kV／cm3组，以1．2kV／cm脉冲强度的兔卵母细胞激活较为理想，脉冲强度增加到2．4kV／cm时，卵细胞死亡率升为67．1％(P＜0．01)。用1．4kV／cm、60μs的直流脉冲电激3次，电脉冲间隔时间分为30、120、240min3组，卵细胞活化率分别为68．6％、91．5％、91．9％（p＜0．01)。     

绵羊卵母细胞的孤雌激活

本文探讨了不同激活方法对绵羊卵母细胞的孤雌激活和其后的发育。结果表明 ,电激活可以激活绵羊卵母细胞孤雌发育到囊胚 ;Ca2 + 载体A2 3187和CHX组合 ,Ionomycin和 6 DMAP组合可以激活绵羊卵母细胞 ,其卵裂率与电激活相比差异显著。 7%乙醇激活绵羊卵母细胞 7min效果较好。不同场强、不同脉冲次数对绵羊卵母细胞激活都有影响 ,以 1.2kV/cm ,间隔 30 μs和 3次脉冲效果较好。而电激活与化学激活联合可以更好的激活绵羊卵母细胞     

不同浓度离子霉素对猪体外成熟卵母细胞孤雌发育的影响

试验探讨了不同浓度的离子霉素结合6-二甲氨基嘌呤激活猪体外成熟卵母细胞后对其孤雌发育的影响。体外成熟卵母细胞分别用10、15、20、25mol/L的离子霉素处理40min,再用2mmol/L的6-DMAP处理8h,各组的卵裂率分别是:(13.11±6.75)%,(22.31±14.18)%,(26.93±24.53)%和(26.72±19.75)%,囊胚率分别是2.26±1.03)%,(2.39±2.41)%,(3.32±3.95)%和(11.76±10.76)%,各组之间差异不显著(P>0.05)。结果表明:用10～25mol/L离子霉素和6-DMAP联合激活猪体外成熟卵母细胞,对其孤雌发育能力有一定影响,但效果不显著;猪卵母细胞采用离子霉素激活的最佳方案是用25mol/L处理40min,再用浓度为2mmol/L的6-DMAP处理8h。     

商会自治的基石——商会自治规范研究

在现代法律秩序中,商会自治规范是制定法的基础和必要的补充,甚至在某些方面替代了制定法;商会自治规范主要包括商会组织规范、行为规范、惩罚规范以及争端解决规范等;其效力仅及于其内部成员;商会自治规范和制定法之间存在冲突,但也存在整合的基础。     

癸氧喹酯干混悬剂含量测定的改进

采用高效液相色谱法测定癸氧喹酯干混悬剂的含量,在2-250μg/mL范围内,峰面积的常用对数与进样量浓度的常用对数呈良好的线性关系,R^2=1(n=5),平均回收率为99.24%~99.51%,RSD在0.05%~0.28%。此方法分析时间短,样品前处理简便、定量结果准确,重现性好,结果满意,为其质量控制提供了依据。     

猪毛色遗传的研究进展

本文概述了猪的毛色类型、猪的毛色遗传模式,着重综述了猪毛色基因分子基础的研究进展,指出存在问题并就未来发展方向做了思考。     

Comparison of 2 methods of centesis of the bursa of the biceps brachii tendon of horses

REASONS FOR PERFORMING STUDY: Centesis of the bicipital bursa using an 8.9 cm long spinal needle has been reported but the alternative of employing a 3.8 cm long hypodermic needle requires validation. OBJECTIVE: To compare the efficacy of 2 different methods of centesis of the bicipital bursa and to evaluate the usefulness of ultrasonographic imaging to determine the location of solution administered when centesis of the bursa is attempted. METHODS: For Trial 1, 6 clinicians, who had no previous experience of centesis of the bicipital bursa, attempted to inject a solution composed of an aqueous radiopaque contrast medium and physiological saline solution (PSS) into the bicipital bursae of 2/12 horses using the previously described distal approach to inject one bursa and a proximal approach to inject the contralateral bursa. The bicipital tendon and bursa were examined ultrasonographically before and after injection; and both shoulders were examined radiographically to identify the location of the medium. In Trial 2, another 6 clinicians, also with no previous experience of centesis, repeated Trial 1, using 6 horses, but the radiopaque contrast medium was mixed with air instead of PSS. RESULTS: Accuracy of centesis using the proximal approach was 39% and that of the distal approach 28%. Ultrasonographic examination of the shoulder allowed the location of solution and air to be accurately predicted in all 12 shoulders examined. CONCLUSIONS: Clinicians who have had no previous experience performing centesis of the bicipital bursa are unlikely to be successful in centesis using either approach. Radiographic examination after injecting a radiopaque contrast medium may be necessary to assess the success of centesis especially if bursal fluid is not obtained during centesis. Injecting air along with the radiopaque contrast medium provides more accurate ultrasonographic confirmation of centesis and better radiographic definition than does injection without air.     

不同样本商品蜂蜜中硒含量的测定

用硝酸和高氯酸消化蜂蜜,使硒游离出来,在微酸性环境下,硒和2,3-二氨基萘(DAN)生成有较强荧光的物质,用环己烷萃取,在激发波长378nm,荧光波长518nm处测定其荧光强度。蜂蜜中硒含量范围:0.10～0.82μg/g。表明:蜂蜜应视为天然富硒营养品。     

乳酸杆菌益生作用机制的研究进展

乳酸杆菌作为益生菌广泛用于人和动物。本文综述了乳酸杆菌改善宿主健康的机制。乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道。文章重点讨论了乳酸杆菌表面成分（表面蛋白、脂磷壁酸和肽聚糖）与肠道受体（Ｃ型凝集素受体、Ｔｏｌｌ样受体和 Ｎｏｄ样受体）,阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制。     

Effects of size of ingestively masticated fragments of plant tissues on kinetics of digestion of NDF

Ingestively masticated fragments were collected and sized via sieving. Different sizes of esophageal masticate and ruminal digesta fragments, and ground fragments of larger masticated pieces were incubated in vitro, and undigested NDF remaining at intervals of up to 168 h of incubation was determined. The ruminal age-dependent time delay (tau) for onset of digestion of NDF was positively correlated (P < 0.004) with the mean sieve aperture estimated to retain 50% of the fragments between successive sieve apertures (MRA). Degradation rate of potentially degradable NDF (PDF) and level of indigestible NDF were not related (P > 0.10) to MRA of masticated and ground fragments. Estimates of tau were positively related to MRA, with slopes of bermudagrass < corn silage < ruminal fragments of corn silage. It was concluded that fragment size-, and consequently, ruminal age-dependent onset of PDF degradation of a mixture of different fragment sizes results in an age-dependent rate of degradation of the more rapidly degrading of two subentities of PDF. Models are proposed that assume a tau before onset of simultaneous degradation of PDF from two pools characterized as having gamma-modeled age-dependency and age-constant rates. The ruminal age-dependent pool seems to be associated with the faster-degrading pool, and its rate parameter increases with range in MRA in the population of fragments. Conceptually, the ruminal age-dependent rate parameter for PDF degradation seems to represent a composite of several effects: 1) effects of the size-dependent tau; 2) range in MRA of the population of ingestively masticated fragments; and 3) subentities of PDF that degrade via more rapid age-dependent rates compared with subentities of PDF that degrade via age-constant rates. The estimated fractional rates of ruminative comminution of ingestively masticated fragments (0.060 to 0.075/h) were of a magnitude similar to the mean fractional rates of PDF digestion (0.030 to 0.085/h), which implies that ruminative comminution may be first-limiting to fractional rate of PDF digestion. The in vivo roles of ingestive and ruminative mastication of fragments on PDF degradation must be considered in any kinetic system for estimating PDF digestion in the rumen. These results and others in the literature suggest that the rate of surface area exposure rather than intrinsic chemical attributes of PDF may be first-limiting to degradation rate of PDF in vivo.     

Mechanism of Action of Probiotic Function of Lactobacilli

乳酸杆菌作为益生菌广泛用于人和动物.本文综述了乳酸杆菌改善宿主健康的机制.乳酸杆菌可通过产生抗菌物质如乳酸、过氧化氢、细菌素,或者通过竞争营养或肠道黏附位点来抑制致病菌;通过诱导黏附素的分泌或阻止细胞凋亡而增强肠道的屏障功能,从而保护肠道.文章重点讨论了乳酸杆菌表面成分(表面蛋白、脂磷壁酸和肽聚糖)与肠道受体(C型凝集素受体、Toll样受体和Nod样受体),阐述了他们结合后启动免疫调节信号,调控肠道免疫功能以发挥改善健康作用的机制.     

Evaluation of serum concentrations of biomarkers of skeletal metabolism and results of radiography as indicators of severity of osteochondrosis in foals

OBJECTIVE: To determine whether serum concentrations of biomarkers of skeletal metabolism can, in conjunction with radiographic evaluation, indicate severity of osteochondrosis in developing horses. ANIMALS: 43 Dutch Warmblood foals with varying severity of osteochondrosis. PROCEDURE: 24 foals were monitored for 5 months and 19 foals were monitored for 11 months. Monthly radiographs of femoropatellar-femorotibial and tibio-tarsal joints were graded for osteochondral abnormalities. Serial blood samples were assayed for 8 cartilage and bone biomarkers. At the end of the monitoring period, foals were examined for macroscopic osteochondrosis lesions. RESULTS: Temporal relationships were evident between certain serum biomarkers and osteochondrosis severity in foals during their first year. Biomarkers of collagen degradation (collagenase-generated neoepitopes of type-II collagen fragments, type-I and -II collagen fragments [COL2-3/4C(short)], and cross-linked telopeptide fragments of type-I collagen) and bone mineralization (osteocalcin) were positive indicators of osteochondrosis severity at 5 months of age. In foals with lesions at 11 months of age, osteochondrosis severity correlated negatively with COL2-3/4C(short) and osteocalcin and positively with C-propeptide of type-II procollagen (CPII), a collagen synthesis marker. Radiographic grading of osteochondrosis lesions significantly correlated with macroscopic osteochondrosis severity score at both ages and was strongest when combined with osteocalcin at 5 months and CPII at 11 months. CONCLUSIONS AND CLINICAL RELEVANCE: The ability of serum biomarkers to indicate osteochondrosis severity appears to depend on stage of disease and is strengthened with radiography. In older foals with more permanent lesions, osteochondrosis severity is significantly related to biomarker concentrations of decreased bone formation and increased cartilage synthesis.