乙醇对绵羊卵母细胞孤雌激活的研究

试验采用乙醇法孤雌激活绵羊体外成熟卵母细胞,研究了卵母细胞体外成熟时间、6-DMAP处理时间、乙醇浓度以及激活时间,对绵羊卵母细胞孤雌激活效果的影响。结果表明:(1)卵母细胞成熟22h、24h和26h卵裂率分别为60.7%、75.2%和74.5%,卵母细胞成熟24h的卵裂率较22h卵裂率,具有明显差异(P<0.05),与26h卵裂率差异不显著(P>0.05);其桑椹胚率38.6%,明显高于26h的桑椹胚率27.3%(P<0.05)。(2)6-DMAP6h的处理组,卵裂率83.4%,较2h卵裂率70.3%,有明显差异(P<0.05),也高于4h卵裂率79.4%,但差异不显著(P>0.05)。(3)乙醇浓度7%,激活3min、5min、7min卵裂率分别为76.7%、81.4%和70.4%,差异均不显著(P>0.05);激活3min囊胚率13.3%,显著高于激活7min囊胚率9.3%(P<0.05)。(4)乙醇浓度7%,激活绵羊卵母细胞5min,卵裂率85.5%,与乙醇浓度为5%的卵裂率72.2%,有显著差异(P<0.05),桑椹胚率和囊胚率,差异分别不显著(P>0.05)。     

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)。     

激活方法对大鼠-小鼠种间体细胞核移植重组胚发育的影响

目的：探讨电激活及化学激活对大鼠-小鼠种间体细胞核移植重组胚发育的影响。方法：采用血清饥饿法同步化处理的SD大鼠成纤维细胞作为供体细胞，常规超排KM小鼠获得卵母细胞，采用盲吸法去核，显微直接注核法构建重组胚，分别采用6-DMAP 乙醇、放线茵酮、透明质酸酶3种化学激活方法和不同电场强度、脉冲电激活重组胚，CZB培养液中培养，观察重组胚发育。结果：电激活：电场强度(V/cm)，脉冲时间(ms)为：1000/100，1500/30，1800/30，2400／40时，卵裂率分别为：17．39％，28．07％，26．52％，0．00％。化学激活：使用透明质酸酶、放线茵酮、6-DMAP 乙醇时卵裂率分别为：5．56％，25．00％和33．91％。结论：电激活和化学激活均可有效激活重组胚：电激活时当电压为1500～1800V/cm、脉冲为30～40ms时，有利于重组胚的发育，电压过高时会将重组胚击碎；本实验所有激活方法中，6-DMAP 乙醇激活法所得卵裂率和桑椹胚率最高。     

不同电融合和激活参数对猪体细胞核移植胚胎发育的影响

目的：摸索猪体细胞核移植最佳的融合和激活条件.方法：利用体外成熟的去核猪卵母细胞为受体,颗粒细胞为供核细胞,透明带间隙注射法构建重构胚,对电融合和激活的参数（电场强度、脉冲宽度、脉冲次数、融合液中Ca^2＋浓度、联合激活参数）进行摸索.结果：发现在电场强度1.5 kV/cm,脉冲宽度30 μs,2次脉冲（间隔3 s）,电融合液中0.1 mmol/L的Ca^2＋浓度下进行电激活,进而用（CB,7.5 μg/ml）＋（6-DMAP,2 mmol/L）激活3.5 h发育率最高,48 h卵裂率为72.5%,第6天桑葚胚率为35.1%;初次尝试了在胚胎培养前48 h加入0.05 mol/L的甘露醇来增加渗透压以减少电激活后重构胚的碎片化程度,但发现卵裂率及桑葚胚发育率与对照组差异不显著.结论：初步得到了适宜的猪体细胞核移植的融合和激活条件,为随后克隆胚早期发育基因的研究打下了基础.     

卵母细胞体外成熟时问对绵羊核移植胚胎发育的影响

将体外成熟的卵母细胞按不同成熟时间进行分组处理，并进行克隆胚胎的生产，实验表明，成熟时间为20-22h并在23-25h完成融合及激活的重构胚其融合率，卵裂率及体外培养到达桑葚胚／囊胚的比例分别为76．2％(176／231)、51．1％(118／231)和21．7％(23／106)，而成熟时间为16～18h及24h以上的卵母细胞进行核移植胚胎的生产，其重构胚各种相应的数据分别为44．9％(84／187)、20．9％(39／187)、10．8％(4／37)以及71．1％(263／367)、48．0％(176／367)、和8．5％(14／176)。     

不同培养体系对猪卵母细胞体外发育潜力的影响

目的:研究猪卵母细胞体外成熟的最佳条件,为用体细胞核移植的方法生产转基因克隆猪提供大量优质的MⅡ期卵母细胞。方法:采用猪卵母细胞体外成熟技术,研究了不同激素组合、猪卵泡液、半胱氨酸和颗粒细胞对猪卵母细胞体外成熟和发育潜力的影响。结果与结论:以15U/mlPMSG 20U/mlhCG成熟率最高,为(70.0±3.30)%,显著高于15U/mlFSH 30U/mlLH、10U/mlPMSG 10U/mlhCG〔成熟率分别为(45.6±5.10)%,(60.0±3.30)%〕,添加激素能促进卵母细胞的分裂,添加15%的猪卵泡液成熟率最高,为(86.7±3.35)%,显著高于5%,10%和20%组〔成熟率分别为(73.3±3.35)%,(71.2±5.08)%和(70.0±6.70)%〕,添加猪卵泡液促进了激活卵母细胞的分裂,在培养基中添加半胱氨酸和颗粒细胞,实验组和对照组成熟率差异不显著(P>0.05),但孤雌激活后卵裂率显著提高(P<0.05)。     

小鼠体外受精、胚胎培养及胚胎快速冷冻的研究

本文系统研究了小鼠受精卵的体内发育与运行规律 ,卵母细胞的体外成熟与受精、单细胞胚胎培养及胚胎快速冷冻 ,结果如下 :①注射hCG后 1 2～ 2 0h受精卵发育至原核期 ,42～ 48h为 2 -细胞期 ,48～ 6 0h为 4-细胞期 ,6 0～ 6 8h为 8-细胞期 ,以上各期受精卵均处于输卵管中 ;75～ 78h为桑椹胚 ,78～ 80h为致密桑椹胚 ,90～92h为早期囊胚 ,92～ 96h为囊胚 ,以上各期均处于子宫角中 ;②培养液中添加激素 ,能显著提高卵母细胞的体外受精率 ,添加FCS和激素组的体外受精率又显著高于单独添加激素组 ,FCS还能显著提高胚胎发育率 ;③在培养液中添加EDTA ,能有效克服小鼠胚胎的 2 -细胞阻断 ,其 2 -细胞胚的发育率达 1 0 0 % ,8-细胞胚的发育率达55%以上 ;牛、羊上皮细胞培养上清也能有效克服 2 -细胞阻断。添加乳酸钠和丙酮酸钠可使 2细胞与 8细胞的发育率显著提高 ;④以D -PBS +甘油 +蔗糖为冷冻液 ,以D -PBS +蔗糖为稀释液 ,对小鼠胚胎进行快速冷冻 ,桑椹胚的存活率为 6 9.3% ,早期囊胚的存活率为 6 0 .4%。以上研究为建立小鼠胚胎库提供了基础     

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...     

选择性单囊胚移植的前瞻性研究

目的 探讨选择性单囊胚移植对体外受精-胚胎移植(IVF-ET)妊娠结局的影响.方法 对海军总医院2007年3月-2011年12月的选择性单囊胚移植进行前瞻性研究.按照移植日胚胎发育阶段以及移植数目分为单囊胚移植组(A组)287例,双囊胚移植组(B组)53例,卵裂期移植组(C组)382例.比较3组患者临床资料、实验室检查结果及妊娠结局的差异;从A、B组中选取最终形成优质囊胚数≥2枚的患者,分为2次单囊胚移植组(n=281)与1次双囊胚移植组(n=53),以累计移植2枚囊胚为例,比较两组单次移植妊娠率和累计妊娠率的差异.结果 与B组相比,A组单次移植周期临床妊娠率下降(P＜0.05),多胎妊娠率降低(P＜0.01).与C组相比,A组临床妊娠率差异无统计学意义(P＞0.05),但种植率明显提高,多胎妊娠率降低(P＜0.01).与B、C组相比,A组卵巢过度刺激综合征(OHSS)的发生率显著降低(P＜0.05).与1次双囊胚移植相比,1次单囊胚移植临床妊娠率下降(P＜0.05),但2次单囊胚移植累计妊娠率与1次双囊胚移植相比差异无统计学意义(P＞0.05).结论 与双囊胚移植和卵裂期移植相比,单囊胚移植可明显降低多胎率、OHSS发生率,适宜有针对性地进行推广应用.     

水牛精子胞质内显微受精技术的初步研究

本研究探讨了水牛精子胞质内显微受精技术的可行性及操作条件的改善对水牛ICSI结果的影响。结果表明 :1.将精子操作液中PVP的浓度从 10 %降低到 5 % ,注射后卵母细胞的存活率无显著差异 (10 0 %vs .97.8% ;P >0 .0 5 ) ,分裂率明显提高 (79.4 %vs.88.6 % ;P <0 .0 5 ) ,囊胚率有所提高 ,但无显著差异 (2 3.5 %vs .2 4 .2 % ;P >0 .0 5 )。 2 .显微注射时卵母细胞第一极体置于相当于时钟 6点的位置与 12点位置相比 ,分裂率无显著差异 (89.5 %vs.86 .2 % ;P >0 .0 5 ) ,但囊胚发育率明显提高 (43.4 %vs.2 7.5 % ;P <0 .0 5 )。 3.离心和不离心的卵母细胞注射后的分裂率分别是 94 .3%和 91.3% ;囊胚发育率分别是 2 3.4 %和 2 7.5 % ,差异均不显著 (P>0 .0 5 )。结论 :1.适当降低PVP的浓度可以促进水牛卵母细胞获得胚胎发育的能力 ,并且不会增加精子显微注射的难度。 2 .注射时水牛卵母细胞的极体位于 6点的位置 ,卵母细胞获得胚胎发育的能力高于 12点位置。3.水牛卵母细胞的低透明度不会影响显微受精的效果。     

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 role of interventional radiology in trauma

Introduction Interventional Radiology has evolved into a specialty having enormous input into the care of the traumatized patient.In all hospitals,regardless of size,the Interventional Radiologist must consider their relationships with the trauma service in order to     

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.     

Diagnóstico ecográfico del neumotórax

The ultrasonographic diagnosis of pneumothorax is based on the analysis of artifacts. It is possible to confirm or rule out pneumothorax by combining the following signs: lung sliding, the A and B lines, and the lung point. One fundamental advantage of lung ultrasonography is its easy access in any critical situation, especially in patients in the intensive care unit. For this reason, chest ultrasonography can be used as an alternative to plain-film X-rays and computed tomography in critical patients and in patients with normal plain films in whom pneumothorax is strongly suspected, as well as to evaluate the extent of the pneumothorax and monitor its evolution.     

Carbohydrate Supplementation during Exercise: Does It Help? How Much is Too Much?

KEY POINTS· Carbohydrate intake during exercise can delay the onset of fatigue and improve performance of prolonged exercise as well as exercise of shorter duration and greater intensity (e.g., continuous exercise lasting about 1h and intermittent high-intensity exercise), but the mechanisms by which performance is improved are different.