发情牛血清和绵羊卵泡液对绵羊卵母细胞体外成熟和受精的影响

本试验研究了绵羊卵泡卵母细胞在体外成熟时,以M-199为基础液,加入FBS,OCS,以及不同直径(小于8mm或大于10mm)卵泡的卵泡液对体外成熟率和体外受精率的影响。在添加同等浓度促性腺激素(FSH,0.4μg/ml;LH,2iu/ml)时,添加5%FBS或5%OCS的成熟率分别为10.00%和69.30%(P<0.05);体外受精分裂率分别为0和60.84%(P<0.05)。在添加以上浓度促性腺激素和5%OCS的基础上添加3%大卵泡液或小卵泡液时,卵母细胞成熟率分别为53.10%和78.20%(P<0.05);体外受精分裂率分别为50.00%和71.67%(P<0.05)。结果表明,发情牛血清配合大卵泡液可以充分满足绵羊卵母细胞体外成熟的需要。     

异种卵泡液对牛卵母细胞体外成熟率的影响

本研究探讨了异种卵泡液对牛卵母细胞体外成熟率的影响。结果表明,在培养基中添加适当浓度的山羊、绵羊、骆驼卵泡液代替血清,牛卵母细胞的成熟率(67．9％、68．5％、65．3％)与血清对照组(72．0％)无显著差异(P＞0．05);在添加山羊、绵羊、骆驼卵泡液的基础上补充激素,卵母细胞的成熟率(70．8％、72．9％、69．7％)也未见明显提高(P＞0．05)。     

不同成熟时间对牛卵母细胞发育及去核效率的影响

本文研究了不同成熟时间对牛卵母细胞的体外成熟、体外受精及后期的体外培养的影响和不同成熟时间的极体出现率。结果表明,牛卵母细胞在成熟16h时极体出现率达69.3%、分裂率为54.7％、囊胚率仅为6.25%;而成熟24h时极体出现率达76.9%、分裂率为70.0%、囊胚率为27.7%,差异均显著（P＜0.05）。成熟16h的卵母细胞刚刚排出第一极体,细胞核与极体非常近,此时去核率较高;而成熟24h的卵母细胞核已远离第一极体。     

三种不同激活方法对牛卵母细胞孤雌激活效果的研究

本试验以体外成熟培养 (IVM ) 2 2～ 2 4h的牛卵母细胞为对象 ,采用以下 3种激活方法 :①电激活 ( 2次直流脉冲 ,时间 30us,强度 1 .8KV/cm ,时间间隔 70us) +环己酰胺 +细胞松弛素B法 ;②电激活 (同上 ) +离子霉素 + 6 -DMAP(二甲基 -氨基嘌呤 )法 ;③ 7%乙醇 +环己酰胺直接激活的方法对体外成熟牛卵母细胞进行孤雌激活实验 ,观察激活后 48h卵母细胞的早期分裂情况。结果表明 ,方法①的卵母细胞分裂率为 31 .32 % ( 2 6 /83) ;方法②的分裂率为 2 7.5% ( 32 /1 1 6 ) ;方法③的分裂率为44.1 1 % ( 45/1 0 2 ) ,与前两种方法相比差异显著 ( p <0 .0 5)。     

小鼠窦前卵泡二维体外培养法的优化研究

目的优化小鼠窦前卵泡二维体外培养体系,探讨卵泡刺激素(FSH)对小鼠窦前卵泡体外发育的影响。方法将分离自14 d雌鼠的初级卵泡(80～100 μm)和早期次级卵泡(110～130 μm)分别培养在浓度为10、100 mU/ml r-FSH的培养液中,观察并记录卵泡体外生长情况和卵母细胞的成熟情况;检测并分析第10天窦卵泡的空间生长情况及不同培养液中卵泡的雌二醇(E₂)分泌水平;Western blotting检测卵泡中FSH受体(FSHR)、类固醇激素合成限速酶3β-羟基类固醇脱氢酶(3β-HSD)、17α-羟化酶(CYP17)及芳香化酶(CYP19)的表达水平。结果初级卵泡在10、100 mU/ml r-FSH培养液中的成腔率(0.00%±0.00%,36.14%±4.02%)及卵母细胞成熟率(0.00%±0.00%,23.54%±7.62%)明显低于早期次级卵泡成腔率(78.63%±4.13%,92.74%±2.54%)及卵母细胞成熟率(48.55%±3.73%,80.88%±4.02%),差异有统计学意义(P<0.05);在100 mU/ml r-FS...     

黄牛体外受精的研究

本实验主要探讨黄牛卵母细胞体外成熟、受精及其受精后胚胎的体外培养。结果表明,卵母细胞在体外培养22小时后:80％以上的卵母细胞发育至中期Ⅱ(MⅡ)阶段。当培养液中添加促卵泡素(FSH,10国际单位/毫升)和雌二醇(E_2 1微克/毫升)时,其培养成熟的卵母细胞不仅受精率提高(83%),而且受精后卵裂率显著提高(67％,P<0.01)。 输卵管细胞单层和细胞上清液不仅能有效地克服牛胚胎的8-细胞阻断,而且对胚胎的发育具有显著的促进作用,二者在培养效果上差异不显著。2—8-细胞阶段胚胎,在输卵管细胞单层、输卵管细胞上清液和对照组(TCM199+10%小牛血清)中培养5天后,分别有77％、74％和27％的早期胚胎发育到桑椹或囊胚。     

牛卵母细胞体外成熟的研究

为了给牛细胞核移植提供卵母细胞 ,1 997年 3月至 2 0 0 1年 3月共进行了 86次卵母细胞体外成熟的实验研究 ,核成熟率 (以出现第一极体为成熟标志 )为 6 5.1 2 %( 2 332 /372 1 )。其中 ,基础培养液为TCM1 99+ 0 .1 2mg/ml丙酮酸钠 + 1 0mMHepes+ 8%NCS + 1 0 μg/mlFSH + 1 0 μg/mlLH + 1 μg/mlE2 (培养液 1 )组进行 6 8批次实验 ,核成熟率为 57.97% ( 1 6 80 /2 898)。基础培养液为TCM 1 99+ 0 .1 2mg/ml丙酮酸+ 8%的发情牛血清 (为 4头发情牛的混合血清 ,培养液 2 )组进行 1 8次实验 ,核成熟率为 79.2 2 % ( 6 52 /82 3)。培养液 1组与培养液 2组之间卵母细胞成熟率差异显著(P <0 .0 5)。培养液 2的组中有 9次添加了 40IU/ml的庆大霉素 ,其核成熟率为72 .1 5% ( 342 /4 74) ;另 9次未添加庆大霉素 ,其核成熟率为 88.83% ( 31 0 /349) ,两者之间无明显差异 (P >0 .0 5)。此外本研究对以去核卵母细胞激活后分裂率作为卵母细胞质成熟的标准作了初步的探讨     

双酚A通过氧化应激干扰小鼠卵母细胞体外成熟

目的 观察双酚A(BPA)对小鼠卵母细胞体外成熟的影响，探讨相关的氧化应激损伤机制。方法 将雌性性成熟ICR小鼠的卵母细胞随机分为对照组、二甲基亚砜(DMSO)组和BPA组，对照组体外正常培养，DMSO组培养液中加入0.1%DMSO,BPA组培养液中加入溶于0.1%DMSO的45?μmol/L BPA。获取处于第二次减数分裂中期(MII)和停滞在生发泡期(GV)的小鼠卵母细胞。采用试剂盒检测MII期和GV期小鼠卵母细胞中活性氧(ROS)的水平；采用免疫荧光法和Western blotting法检测MII期和GV期小鼠卵母细胞中抗氧化酶的表达水平。结果 与对照组比较，体外成熟过程中接触45?μmol/L BPA的BPA组小鼠卵母细胞成熟率明显降低(26.32%±1.12%vs. 98.22%±0.89%,P<0.05),GV期和MII期卵母细胞中ROS水平明显升高(P<0.05)，过氧化氢酶(CAT)和超氧化物歧化酶2(SOD2)水平明显降低(P<0.05)。结论 小鼠卵母细胞体外接触45?μmol/L BPA可增高ROS水平，抑制抗氧化酶CAT和SOD2的表达，引起氧...     

猪卵母细胞电激活参数的研究

以体外成熟的猪卵母细胞为对象 ,研究不同卵龄、电场强度、脉冲次数和激活液Ca 、Mg 对猪卵母细胞电激活效果的影响。结果表明 ,1次脉冲就足以激活猪卵母细胞 ,电场强度以 1 2 50～ 1 50 0V/cm为优 ,含Ca 、Mg 的激活液激活效果显著高于非电解质液 ,卵龄以 54h为宜     

猪卵子体外受精研究

本研究旨在探讨影响猪卵子体外成熟、猪精子体外获能和猪卵子体外受精的某些因素。主要研究:(1)猪卵体外成熟所需的时间;(2)促性腺激素和猪卵泡液对猪卵体外成熟的影响;(3)咖啡因、肝素和离子钙I-A_(23187)对猪精子体外获能的促进作用;(4)不同体外受精次数对猪卵体外受精结果的影响;(5)不同种类猪精液的体外受精能力。各试验组的结果比较主要根据体外受精后卵子的卵裂率,数据用卡方(x~2)进行统计学分析。 试验结果表明:(1)猪卵子体外成熟需要32—36小时,经32—36小时体外成熟的 卵子体外受精后卵裂率(2—4细胞期)达31.16％,明显高于其他对照组(P<0.01);(2)促性腺激素和猪卵泡液能够促进猪卵子体外成熟,当0.25国际单位/毫升PMSG和10％猪卵泡液,或者0.25国际单位/毫升FSH和10％猪卵泡液分别加入到猪卵子体外成熟中,经此培养后的猪卵子体外受精卵裂率分别达到23.95％和35.00％,明显高于对照组(P<0.05);(3)咖啡因、肝素、离子钙I-A_(23187)对猪精子体外获能有促进作用。在精子预培养液中加入0.05毫克/毫升肝索,在受精液中加入2毫克/毫升咖啡因,获得了体外受精后最高的卵裂率36.16％,明显高于其他组(P<0.01);(4)多次体外受精法有助于提高体外受精率,因为卵子的体外成熟是不同步的,但是超过2次又是有害的。用2     

Knee injury and Osteoarthritis Outcome Score (KOOS) - validation of a Swedish version

The Knee injury and Osteoarthritis Outcome Score (KOOS) is a self-administered instrument measuring outcome after knee injury at impairment, disability, and handicap level in five subscales. Reliability, validity, and responsiveness of a Swedish version was assessed in 142 patients who underwent arthroscopy because of injury to the menisci, anterior cruciate ligament, or cartilage of the knee. The clinimetric properties were found to be good and comparable to the American version of the KOOS. Comparison to the Short Form-36 and the Lysholm knee scoring scale revealed expected correlations and construct validity. Item by item, symptoms and functional limitations were compared between diagnostic groups. High responsiveness was found three months after arthroscopic partial meniscectomy for all subscales but Activities of Daily Living.     

Endovascular management of carotid-cavernous fistulas

Objective To investigate endovascular treatment of traumatic direct carotid-cavernous fistulas （CCF） and their complications such as pseudoaneurysms. Methods： Over a five-year period, 22 patients with traumatic direct CCFs were treated endovascularly in our institution. Thirteen patients were treated once with the result of CCF occluded, 8 twice and 1 three times. Treatment modalities included balloon occlusion of the CCF, sacrifice of the ipsilateral internal carotid artery with detachable balloon, coll embolization of the cavernous sinus and secondary pseudoaneurysms, and covered-stem management of the pseudoaneurysms. Results All the direct CCFs were successfully managed endovascularly. Four patients developed a pseudoaneurysm after the occlusion of the CCF with an incidence of pseudoaneurysm formation of 18.2% （4/22）. A total number of 8 patients experienced permanent occlusion of the ICA with a rate of ICA occlusion reaching 36.4% （8/22）. Followed up through telephone consultation from 6 months to 5 years, all did well with no recurrence of CCF symptoms and signs. Conclusion Traumatic direct CCFs can be successfully managed with endovascular means. The pseudoaneurysms secondary to the occlusion of the CCFs can be occluded with stent-assisted coiling and implantation of covered stents.     

The acutely limping child

Acute limping may be the result of multiple pathologies in children. The differential diagnosis varies based on the age of the child. Irrespective of age, the initial imaging work-up includes AP and frog leg radiographs of the pelvis and ultrasound; MRI may sometimes be helpful. In children less than 3 years, infections and trauma are most frequent. MRI is the imaging modality of choice when osteomyelitis is clinically suspected. Between the ages of 3 and 10 years, transient synovitis of the hip and Legg-Calvé-Perthes disease are main considerations but infection, inflammation and focal bony lesions are also considered. In children over 10 years, slipped capital femoral epiphysis also is considered.     

Do Balance Board Training Programs Reduce the Risk of Ankle Sprains in Athletes?

Introduction Ankle sprains are the most common musculo-skeletal injury that occurs in athletes,particularly in sports that require jumping and landing on one foot such as soccer,and basketball(1-4).These injuries often result in significant time loss from participation,long-term disability,and have a major impact on health care costs and resources(5-8).     

High Intensity Interval Training:New Insights

KEY POINTS ·High-intensity interval training(HIT)is characterized by repeated sessions of relatively brief，intermittent exercise.often performed with an“a11 out”effort or at an intensity close to that which elicits peak oxygen uptake(i.e.，≥90%of VO2 peak).     

Developmental validation of the PowerPlex(®) ESI 16 and PowerPlex(®) ESI 17 Systems: STR multiplexes for the new European standard

In response to the ENFSI and EDNAP groups’ call for new STR multiplexes for Europe, Promega^® developed a suite of four new DNA profiling kits. This paper describes the developmental validation study performed on the PowerPlex^® ESI 16 (European Standard Investigator 16) and the PowerPlex^® ESI 17 Systems. The PowerPlex^® ESI 16 System combines the 11 loci compatible with the UK National DNA Database^®, contained within the AmpFlSTR^® SGM Plus^® PCR Amplification Kit, with five additional loci: D2S441, D10S1248, D22S1045, D1S1656 and D12S391. The multiplex was designed to reduce the amplicon size of the loci found in the AmpFlSTR^® SGM Plus^® kit. This design facilitates increased robustness and amplification success for the loci used in the national DNA databases created in many countries, when analyzing degraded DNA samples. The PowerPlex^® ESI 17 System amplifies the same loci as the PowerPlex^® ESI 16 System, but with the addition of a primer pair for the SE33 locus. Tests were designed to address the developmental validation guidelines issued by the Scientific Working Group on DNA Analysis Methods (SWGDAM), and those of the DNA Advisory Board (DAB). Samples processed include DNA mixtures, PCR reactions spiked with inhibitors, a sensitivity series, and 306 United Kingdom donor samples to determine concordance with data generated with the AmpFlSTR^® SGM Plus^® kit. Allele frequencies from 242 white Caucasian samples collected in the United Kingdom are also presented. The PowerPlex^® ESI 16 and ESI 17 Systems are robust and sensitive tools, suitable for the analysis of forensic DNA samples. Full profiles were routinely observed with 62.5 pg of a fully heterozygous single source DNA template. This high level of sensitivity was found to impact on mixture analyses, where 54–86% of unique minor contributor alleles were routinely observed in a 1:19 mixture ratio. Improved sensitivity combined with the robustness afforded by smaller amplicons has substantially improved the quantity of data obtained from degraded samples, and the improved chemistry confers exceptional tolerance to high levels of laboratory prepared inhibitors.