猪精液0.5 ml细管快速冷冻和解冻方法的优化

 【目的】优化冷冻-解冻方法,提高0.5 ml细管快速冷冻保存猪精液的质量。【方法】冷冻方法：分别将细管水平置于液氮面上方3、5、7、9和11 cm处,每一水平位置细管分别熏蒸3、5、10、15和20 min后投入液氮保存,37℃×30 s水浴解冻,确定最优冷冻方法;在选择最优冷冻方法的基础上,分别采用37℃×30s(对照组)、42℃×25 s、47℃×20 s、52℃×15 s、57℃×10 s和62℃×5 s共6种方法水浴解冻,确定最佳解冻方法。【结果】采用3 cm×10 min冷冻方法,解冻0、8 h精液中精子活率、质膜完整率和顶体完整率均最高。采用相对“高温短时”解冻,精子活率、质膜完整率、顶体完整率和线粒体膜电位均较高,同时丙二醛浓度降低;57℃×10 s和62℃×5 s解冻的精子各项指标无显著差异（P＞0.05）,但前4项指标显著高于对照组（P＜0.05）,丙二醛浓度显著降低（P＜0.05）。【结论】优化猪精液0.5 ml细管快速冷冻的冷冻和解冻方法可以显著提高解冻质量,采用3 cm×10 min冷冻、57℃×10 s或62℃×5 s解冻方法效果最佳。     

α-Amylase inhibitor in local Himalyan collections of Colocasia: Isolation, purification, characterization and selectivity towards α-amylases from various sources

The α-amylase inhibitor from corms of Colocasia collected from Bhota village of Hamirpur district, Himachal Pradesh was purified to 17.21 folds with 61.61% recovery using ammonium sulfate precipitation, gel filtration chromatography (sephadex G-200) and ion exchange chromatography (DEAE-sephadex). A single band of the purified inhibitor was obtained by Native-PAGE. SDS-PAGE revealed the purified inhibitor to be a monomer with molecular weight of 13,900 daltons. The nature of inhibition was found to be of non-competitive type as determined by Lineweaver-Burk plot and a K_i value of 0.54 nmole was obtained by Dixon’s plot. The inhibitor was found to be heat stable and retained 81.50% activity at 70 °C temperature. Inhibitor was found to have pH optima of 6.9. The purified inhibitor was found to have inhibitory activity against α-amylases extracted from the larvae of Callosobruchus chinensis, Tribolium castaneum, Corcyra cephalonica and midgut α-amylase of Spodoptera littoralis. 100% larval mortality of C. cephalonica was observed when fed on wheat flour mixed with 0.0036% (w/w) of purified inhibitor. Purified α-amylase inhibitor was found to inhibit the activity of human salivary α-amylase. It also had inhibitory activity against potato α-amylases and reduced sugar content in treated potato slices. The purified inhibitor was found to be a glycoprotein. In the present study, the ability of the inhibitor to inhibit insect amylases highlights its possible role in pest resistance and post harvest decay of crop plants. Inhibitory activity of α-amylase inhibitor against mammalian amylases could suggest its potential in treatment of diabetes and cure of nutritional problems, which result in obesity.     

250g/L嘧菌酯悬浮剂防治武陵山区马铃薯及番茄晚疫病的试验研究

进行250 g/L嘧菌酯悬浮剂防治武陵山区马铃薯及番茄晚疫病的试验研究,结果表明:250 g/L嘧菌酯悬浮剂茎叶处理对马铃薯及番茄晚疫病均有一定的防效,速效性及持效性均显著或极显著优于对照药剂,并对作物安全,防病增产效果突出。     

Immunological properties of porcine enterotoxigenic Escherichia Coli heat-stable (ST) enterotoxins

Cross-linking semipurified and purified heat-stable enterotoxin (2023 ST) to carrier proteins with glutaraldehyde pigs and rabbits were immunized. With ST antisera thus raised, a relatively simple, two-step method for isolation of ST is described. The method involved immunoadsorbent column chromatography procedure, and elution of the retained material with 6 M urea at pH 3.0 yielded purified ST with enterotoxin activity, controlled in suckling mouse test. ST purity was checked with a special staining technique in PAGE, anionic-exchange chromatography and immunodiffusion.     

后备奶牛痒螨病的诊治

牛痒螨病是一种由痒螨属中的痒螨引起的慢性接触性皮肤外寄生虫病。易引起寄生宿主不安，剧烈瘙痒，皮肤创伤，形成痂皮。影响奶牛采食、休息，从而影响奶牛生长发育和产奶量。     

几种SDS—PAGE蛋白质染色方法的比较

本文在对已报道的三种不同的SDS—PAGE蛋白质染色方法加以改进的基础上,对各测定方法的灵敏度、重复性及有关影响染色的因素进行了比较和讨论。结果表明,改良的Pharmacia公司银染色法所得到的结果最好,适合作为实验室常规检测手段。     

几种测定灰树花多糖中蛋白质含量方法的比较研究

用双缩脲法、紫外吸收法、考马斯亮蓝G－250染色法三种方法对灰树花多糖中蛋白质含量进行测定，并同微量凯氏定氮法进行对比。结果表明：在蛋白质量浓度0．1-1.0mg/mL范围内，其测量误差分别为双缩脲法大于50％，紫外吸收法大于30％，考马斯亮蓝G－250染色法最低可小于3％，考马斯亮蓝G－250染色法简单，迅速，且相对较为准确，是测定灰树花多糖中蛋白质含量的有效方法。     

Determination of the concentration of protein in cerebrospinal fluid with a new dye-binding method

The protein concentration of cerebrospinal fluid (CSF) was measured on 50 samples with a new dye-binding method using Coomassie Brilliant Blue G-250 (CBB). The results of this method correlated well (r = 0.99) with those of the trichloroacetic acid-Ponceau S method (TCA-PS). Since the CBC method involves only one step, it is recommended as a simple method of determining protein concentration in CSF.     

Current development and use of hemoglobin-based oxygen-carrying (HBOC) solutions

Objective: To review the human and veterinary literature on the current development and use of hemoglobin‐based oxygen‐carrying solutions. Human data synthesis: Hemoglobin‐based oxygen‐carrying (HBOC) solutions have been developed extensively over the last 3 decades. Early problems associated with pure hemoglobin and cytoskeleton residues have been resolved with chemical modification of the hemoglobin tetramer resulting in effective oxygen‐carrying molecules of either human or bovine origin. The limited availability of human red blood cells and concerns of disease transmission, the difficulty in mass production of genetically produced hemoglobin solutions (recombinant hemoglobin), and the wide availability of bovine blood have resulted in the development of bovine‐derived HBOC solutions. Research efforts have been directed toward determining the effects of HBOC solutions on tissue perfusion as the target uses of HBOC solutions in human medicine are the perioperative period, shock, and trauma fluid resuscitation. The most controversial issues regarding the cardiovascular effects of HBOC solutions surround increased vasoactivity. Some HBOC formulations have been removed from advanced clinical trials due to intense vasoactivity resulting in increased morbidity. There are currently 3 HBOC solutions in the latter stages of phase III clinical trials: Hemolink®, ^a Hemopure®, ^b and PolyHeme®. ^c The hemoglobin source of Hemopure® is bovine, and the hemoglobin source for Hemolink® and Polyheme® is human. Veterinary data synthesis: The only HBOC solution that has gained approval from the FDA is the veterinary product Oxyglobin®. ^d Oxyglobin® is 13 g/dL of ultrapurified, polymerized hemoglobin solution of bovine origin in a modified lactated Ringer's solution. There is a significant colloid effect and it also provides a plasma source of oxygen‐carrying capacity. The solution is stable at room temperature for 3 years; there is no special preparation required prior to use and no cross‐match is required prior to administration (contains no cell membranes). Veterinary publications on the use of Oxyglobin® include laboratory investigations in dogs, cats, and horses for use as a resuscitation fluid and for the treatment of anemia. Clinical use of Oxyglobin® in dogs, cats, birds, horses, and other mammalian species has been reported in several publications. Conclusion: The search for a safe, effective HBOC solution for use in human medicine is ongoing. Soon, there will be one or several products approved for use in the perioperative period and/or for the treatment of shock and trauma. The practice of veterinary emergency and critical care has been provided a unique opportunity to apply the use of an HBOC solution (Oxyglobin®) to various aspects of perfusion and oxygen‐carrying needs. Continued clinical experience and research is essential in understanding the use of HBOC solutions in veterinary medicine.     

蛋白质电泳中考马斯亮蓝R 250染色方法的优化与改进

为了优化与改进蛋白质电泳中传统考马斯亮蓝（Coomassie brilliant blue,CBB）R 250染色法凝胶背景深、脱色时间长、试剂消耗多等弊端,通过优化试验,建立了一种简单经济的 CBB R 250染色方法。与传统方法相比,该方法在染色液配制中降低了甲醇、冰醋酸的比例,增加了硫酸铵成分,从而有效改善了染色效果,改进后的方法,染色几乎无背景,不需专用脱色液,只需用清水洗涤即可观察效果,大大缩短了试验时间,节约了成本,值得推广应用。