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)。     

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     

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)。     

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)。     

卵子发生过程中生殖细胞与体细胞的相互作用

引言 雌性哺乳动物在胎儿晚期或生后不久,初级卵母细胞开始第一次成熟分裂,然后停止前期的双线期,亦称核网期。核网期随种类不同可持续数周至数年。核网期的卵母细胞由一层体细胞(颗粒细胞的前身)包被,组成原始卵泡。一组处于双线期的卵母细胞进入生长阶段,其时间长短随动物种类而异。生长期末,卵母细胞有恢复成熟分裂的能力,但只有在内源性促性腺激素峰之后才     

牛卵母细胞体外成熟培养时间对体外受精及早期胚胎体外发育的影响

本文报道了牛卵母细胞经不同时间体外成熟培养对其后体外受精及早期胚胎体外发育能力的影响。 实验用的母牛卵巢从附近的屠宰场收集。卵母细胞采用穿刺抽取法采集。将外观形态正常的卵母细胞随机分成6组,在含10％发情牛血清的TCM-199培养液中分别经12(G1)、16(G2)、20(G3)、24(G4、对照组)、28(G5)和32小时(G6)的体外成熟培养(IVM)后进行体外受精(IVF),早期胚胎(2-8细胞阶段)在采用卵泡颗粒细胞作为支持细胞的单层上皮细胞液滴中进行 复合培养至囊胚及孵化囊胚阶段。结果表明,经过12—32小时IVM的牛卵母细胞,在IVF后48小时检查,卵裂率均可达到81％以上(80.9％—91.9％);但IVM时间短至12小时(G1)及超过24小时(G5、G6)的卵裂率(分别为80.9％,83.5%和83.4％)显著地低于24小时的对照组(G4,91.9%,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。     

应用激素调控绵羊繁殖生产之初探

为探索应用激素调控绵羊繁殖生产,诱导绵羊非繁殖季节发情配种,挑选细毛羊和部分土种羊经产母羊共计257只,分成三组,即Ⅰ组70只细毛羊,第Ⅱ组157只细毛羊和Ⅲ组30只土种羊,用外源促性腺激素进行诱导发情处理,结果Ⅰ组产羔率达到了51.43%,比Ⅱ组的产羔率32.48%高出18.95个百分点(P<0.01),比Ⅲ组的6.67%高出44.76个百分点(P<0.01),Ⅱ组比Ⅲ组高出25.81个百分点(P<0.01)。研究表明应用生殖激素对细毛羊在非繁殖季节实施诱导发情是可行的,土种羊对外源促性腺激素的应答没有细毛羊敏感。     

西门塔尔牛体外胚胎生产效率分析

【目的】文章旨在分析西门塔尔牛活体采卵-体外受精（OPU-IVF）体外胚胎生产效率的影响因素,为提高西门塔尔牛体外胚胎生产效率、优化西门塔尔牛体外胚胎生产流程提供生产依据。【方法】本实验以未经激素处理的西门塔尔牛卵丘卵母细胞复合体（COCs）、进口肉牛冻精为实验材料,通过卵母细胞活体采集（OPU）、体外成熟（IVM）、体外受精（IVF）、早期胚胎体外培养（IVC）等技术手段,对体外受精后第7 d囊胚率进行统计分析。【结果】不同公牛精液对囊胚发育有影响,不同供体母牛对囊胚发育率影响显著。【结论】相同卵母细胞利用率水平的供体母牛,回收COCs体外受精后,囊胚率与冻精有关（P<0.05）;不同卵母细胞利用率水平的供体母牛,回收COCs体外受精后,囊胚率随卵母细胞利用率升高而升高（P<0.05）。     

血清与卵泡液中血管内皮生长因子在控制性超排卵周期中的变化

目的 本研究通过观察控制性超排卵患者的血清及卵泡液中血管内皮生长因子的变化,探讨血管内皮生长因子与卵泡发育的关系。方法 50个行体外受精-胚胎移植治疗周期的患者为研究对象。分别于月经周期第2天、取卵日抽取静脉血20例,收集取卵日卵泡直径≥18mm或≤16mm卵泡的卵泡液各25例,用酶联免疫吸附试验方法测定VEGF的浓度。结果 取卵日血清VEGF水平(116.9±17.1)mg/L较月经第2天的(92.0±17.2)ng/L明显增高(P<0.001);取卵日卵泡直径≥18mm卵泡液中VEGF水平(1630.6±144.8)ug/L较卵泡直径≤16mm(1475.9±155.5)ng/L高(P<0.05);20例取卵日卵泡液水平(1492.8±184.9)ng/L显著高于同期血清的(116.9±17.1)ng/L(P<0.001)。结论 控制性超排卵周期中血清及卵泡液中VEGF水平均升高,可能参与卵泡发育、卵母细胞成熟。     

Glutamine supplementation in vitro and in vivo,in exercise and in immunodepression

In situations of stress, such as clinical trauma, starvation or prolonged, strenuous exercise, the concentration of glutamine in the blood is decreased, often substantially. In endurance athletes this decrease occurs concomitantly with relatively transient immunodepression. Glutamine is used as a fuel by some cells of the immune system. Provision of glutamine or a glutamine precursor, such as branched chain amino acids, has been seen to have a beneficial effect on gut function, on morbidity and mortality, and on some aspects of immune cell function in clinical studies. It has also been seen to decrease the self-reported incidence of illness in endurance athletes. So far, there is no firm evidence as to precisely which aspect of the immune system is affected by glutamine feeding during the transient immunodepression that occurs after prolonged, strenuous exercise. However, there is increasing evidence that neutrophils may be implicated. Other aspects of glutamine and glutamine supplementation are also addressed.     

糖尿病鼠血清及睾丸中一氧化氮含量和一氧化氮合酶活性的变化

目的研究1型糖尿病大鼠血清和睾丸中一氧化氮(NO)含量和一氧化氮合酶(NOS)活性的动态变化。方法63只Wistar雄性大鼠随机分为对照组(C)和糖尿病组(D)，采用四氧嘧啶腹腔注射复制糖尿病动物模型。糖尿病组分别于观察1、2、3、4、5、6、7和8周后断头处死动物，取血收集血清，取睾丸制备组织匀浆，分别测定NO含量和NOS活性。结果与对照组比较，糖尿病组血清NO含量升高，NOS活性先升后降；睾丸组织在诱模后NO含量下降，而NOS活性变化不明显，随后两者均呈升高趋势。结论糖尿病大鼠血清和睾丸NO含量和NOS和活性均有改变。     

2018年三亚雪古丽牛羊布鲁氏菌病及口蹄疫监测与风险评估

为持续性确保三亚雪古丽牛羊口蹄疫无疫及处于布鲁氏菌净化状态,雪古丽对自有及引进羊群进行了口蹄疫免疫抗体检测及病原学检测,同时对羊群进行了布鲁氏菌病抗体检测.口蹄疫免疫抗体检测应用液相阻断ELISA方法;口蹄疫病原学检测采用多重RT-PCR;布鲁氏菌病用虎红平板方法检测.检测结果:未检测到布鲁氏菌抗体阳性动物,牛羊中目前不存在口蹄疫病畜或健康带毒畜,牛羊口蹄疫免疫抗体未达到国家规定水平的应及时补免.2018年三亚雪古丽牛羊布鲁氏菌病及口蹄疫监测结果表明,雪古丽的羊场目前不存在布鲁氏菌感染情况;牛羊中仅牛O型口蹄疫群体免疫水平达到国家重大动物疫病规定标准,免疫抗体不合格动物应及时给予补免,病原学检测结果表明目前雪古丽的牛羊及引进羊未被口蹄疫病毒感染.     

Icariin protects against radiation-induced mortality and damage in vitro and in vivo

Abstract

Purpose: The present study aimed to investigate the potential protective effects of icariin both in vivo and in vitro, an active flavonoid glucoside derived from medicinal herb Epimedium, and its possible mechanisms against radiation-induced injury.

Methods: Male C57BL/6 mice were exposed to lethal dose (7?Gy) or sub-lethal dose (4?Gy) of whole body radiation by X-ray at a dose rate of ～0.55?Gy/min, and icariin was given three times at 24?h and 30?min before and 24?h after the irradiation. After irradiation, hematological, biochemical, and histological evaluations were performed. We further determined the effect of icariin on radiation-induced cytotoxicity and changes in apoptosis-related protein expression.

Results: Icariin enhanced the 30-day survival rates (20 and 40?mg/kg) in a dose-dependent manner, and protected the radiosensitive organs such as intestine and testis from the radiation damages. Moreover, hematopoietic damage by radiation was significantly decreased in icariin-treated mice as demonstrated by the increases in number of peripheral blood cells, bone marrow cells (1.7-fold), and spleen colony forming units (1.7-fold). In addition, icariin decreased the radiation-induced oxidative stress by modulating endogenous antioxidant levels. Subsequent in vitro studies showed that icariin effectively increased cell viability (1.4-fold) and suppressed the expression of apoptosis-related proteins after irradiation.

Conclusion: These results suggest that icariin has significant protective effects against radiation-induced damages partly through its anti-oxidative and anti-apoptotic properties.     

西宁地区新生儿缺氧缺血性脑病与肾功能损害探讨

目的：探讨高海拔地区新生儿HIE与肾功能损害的关系。方法：对116例HIE足月儿和88例足月儿正常新生儿做血尿素氮（Bun）及肌酐（Cr）检测,并进行分析观察。结果：116例HIE患儿血BuN〉7．14mmol／L96例,血肌酐〉88．4mmol／L91例。HIE组血Bun、Cr显著高于正常对照组（P〈0．01）,中度HIE组Bun、Cr显著高于轻度HIE组（P〈0．05）,重度HIE组Bun、Cr显著高于中度HIE（P〈0.05）。结论：新生儿窒息缺氧会导致脑损伤和肾功能损害,血Bun、Cr可做为判断肾功能受损指标,而且窒息程度越重,肾功能损害越严重。     

The stability and distribution of Tc-HIDA in vivo and in vitro

Tc-HIDA is a promising new agent for imaging the biliary system. In this study radiochromatography on paper or Sephadex G25 gel has been used to measure the proportions of TcHIDA, hydrolysed Tc and pertechnetate in solutions from a comercial kit (CIS TCK 15) and in body fluids from patients and rat receiving this radiopharmaceutical. The tissue distribution in male and female rats has shown the radiopharmaceutical to be rapidly removed from the blood by the liver and then excreted via the intestines. There appears to be a sex difference in the uptake in the kidneys and in the urinary excretion in both rats and patients.     

Measurement of GABA and contaminants in gray and white matter in human brain in vivo.

A preliminary study of discrimination between GABA and macromolecules (MMs) in human brain by proton double quantum filtering (DQF) at 3.0 T in vivo is presented. GABA-tuned and MM-tuned DQ filters were designed with dual-band 180 degrees radiofrequency (RF) pulses that were tuned for selective refocusing of GABA (3.0 and 1.9 ppm) and putative MM resonances (3.0 and 1.7 ppm), respectively. GABA and putative MM signals were extracted from a combined analysis of the filtered mixture signals and the calculated editing yields. Unexpectedly, the GABA and putative MM signals exhibited a similar doublet linewidth at the optimized TE = 82 ms. Furthermore, substantial MM-tuned DQF signal remained at TE = 148 ms, indicating the presence of a component other than MM. With water segmentation data, the GABA-tuned and MM-tuned DQF measures from the medial prefrontal and left frontal lobes were combined to give the concentrations of GABA and the additional component as 1.1 +/- 0.1 and 0.8 +/- 0.1 mM (mean +/- SD, N=3) for gray matter (GM) and 0.4 +/- 0.1 and 0.7+/-0.1 mM (N=3) for white matter (WM), respectively.     

The stability and distribution of Tc-HIDA in vivo and in vitro

Tc-HIDA is a promising new agent for imaging the biliary system. In this study radiochromatography on paper or Sephadex G25 gel has been used to measure the proportions of TcHIDA, 'hydrolysed Tc' and pertechnetate in solutions from a commercial kit (CIS TCK 15) and in body fluids from patients and rats receiving this radiopharmaceutical. The tissue distribution in male and female rats has shown the radiopharmaceutical to be rapidly removed from the blood by the liver and then excreted via the intestines. There appears to be a sex difference in the uptake in the kidneys and in the urinary excretion in both rats and patients.