Immunohistochemical Study of Hexose Transporters GLUT-2 and GLUT-5 in Birds Gastrointestinal Tract
免疫组化的原理与操作
化技术成为当今生物医学中形态、功 能代谢综合 研究的一项有力工具。其应用范围深达医学各个 学科，是目前生命科学工作者应该掌握的基本技 术之一； ——国内起步晚，从70s开始。
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二、IHC的现状和发展前景
（一）方法学
如将该法中的卵白素（avidin）变换成链霉卵白素(streptavidin)，LAB法即成LsAB法，或称SP法。
（B ★为生物素标记的过氧化物酶）
AB★C复合物是avidin与酶联biotin 在使用前30min 配置，混匀。1个avidin分子结合了3个biotin分子，剩下 1个结合点与2抗的生物素结合，avidin起桥联作用。
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（1）ABC复合物的制备：
（2）ABC法反应原理：
特点：ABC法敏感性更高，比PAP法敏感20~40倍，背景 也更清晰。
特点或优点：（1）特异性强、灵敏度高；
（2）可定性、定位、定量观察；
（3）将形态学改变与功能和代谢变化结
合起来；
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IHC不仅具有传统形态学（包括LM和EM水平） 能对组织和细胞进行仔细、客观观察的优点（特别 是经苏木精或伊红复染后），而且还克服了传统免 疫学反应只能定性和定量（离体、液相），而不能 定位的缺点。IHC则可在原位和固相对染色结果进 行观察。目前IHC的定位可精确到亚微结构水平。
荧光素直接标记特异性抗体（一抗）上，标记抗 体与抗原结合（在切片上）荧光显微镜下观察→ 检测抗原。
优点：简单，时间短， 特异性强。
缺点：灵敏度低，所 需抗体量大 （不经济）。
应用：基本不用了！！
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Science：英国科学家揭示神经干细胞重回静默状态的重要机制
Science：英国科学家揭示神经干细胞重回静默状态的重要机制2016年7月18日讯/生物谷BIOON/ --近日，著名国际学术期刊Science刊登了英国弗朗西斯克里克研究所-米尔希尔实验室研究人员的一项最新研究进展，在这篇文章中他们发现了增殖状态下的神经干细胞如何重新回到静默状态，从而维持神经干细胞池的平衡。
海马体是大脑中负责调节记忆和情绪的重要区域，其中的神经干细胞能够产生新的神经元，即使在成年阶段也具有这样的能力。
形成多少个新的神经元以及何时形成新的神经元取决于神经干细胞池的静默和增殖之间的平衡。
那么什么样的信号能够让处于增殖状态的神经干细胞回到静默状态引起了研究人员的兴趣。
研究人员发现一个促进细胞增殖的关键转录因子发生泛素化降解，就能调节神经干细胞回到静默状态，但是这种静默状态与神经干细胞的原始状态并不相同，这种处于静默但又活化状态的神经干细胞维持了神经干细胞池的平衡。
研究人员表示，神经干细胞的静默对于成体神经干细胞的长期维持非常重要，微环境信号能够调节神经干细胞从静默状态变成激活状态，但是增殖状态的神经干细胞如何重新回到静默状态仍未可知。
在这项研究中，研究人员发现E3连接酶Huwe1是增殖状态下的成年小鼠海马体神经干细胞回到静默状态的一个必要分子。
Huwe1能够通过泛素化降解系统使增殖状态下的海马体神经干细胞中促进细胞增殖的转录因子Ascl1变得不稳定，进而阻止细胞周期蛋白D的积累促进增殖细胞回到静默状态。
当神经干细胞回到静默状态，增殖的神经干细胞池会逐渐耗竭。
研究人员认为长期维持海马体神经元生成需要依赖于快速降解这种关键的促激活因子使神经干细胞回到一种暂时性的静默状态。
这项研究发现了促进增殖的神经干细胞回到静默状态的一条重要分子机制，机体通过这种静默状态和增殖状态的神经干细胞池平衡维持正常的神经元生成，该研究对于神经疾病研究和治疗甚至癌症研究都有重要提示意义。
免 疫 组 织 化 学
核酸分子探针-杂交-免疫组织化学 放大和显示杂交信号 杂交免疫组织化学
基因重组技术 免疫组织化学 图像分析
单抗技术
技术
流式细胞术
激光共聚焦显微术
免疫组织化学的全过程： 抗原的提取和纯化 免疫动物或细胞交融〔单克隆抗体〕 抗体效价检测和提取、纯化 标志抗体 细胞、组织切片标本的制备 免疫组织化学反响和显色 察看和记录结果
● 切片边缘、刀痕或皱折区域 、坏 死或挤压的细胞区、胶原结缔组织等， 常表现为一样的阳性染色强度，不能 用于判别阳性。
第三章 免疫组织化学常用方法引见
第一节 免疫酶细胞化学
免疫酶细胞化学是免疫组织化学中最常用的方法，它 是在抗原抗体特异反响存在的前提条件下，借助于酶细胞化 学手段，检测某种物质〔抗原／抗体〕在组织细胞内的存在 部位。即预先将抗体与酶连结〔酶标抗体〕，再使其与组织 内特异性抗原反响，经细胞化学染色后，在光镜或电镜下察 看分析。
后再用显微镜察看〔免疫酶〕
1. 加强特异性染色的方法 ● 蛋白酶消化法 胃蛋白酶、胰蛋白酶 ● 适宜的抗体稀释度 第一抗体 ● 温育时间 37℃ 30-60 min, 4℃ 过夜 ● 多层染色法〔双层、三层〕
2. 减少或消除非特异性染色的方法 加二抗 显色剂的浓度、温育时间
二抗 1
一抗
1:50
1:100
1:200
1:400
_____________________________________ ___________
1:50
+++
+++
◆ 以中等阳性稀释度为佳 ◆ 抗体稀释液的配制
0.01mol/L pH7.4 PBS or TBS
chinadaily双语新闻：挑战动物起源理论丹麦海绵需氧极少
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One of science's strongest dogmas is that complex life on Earth could only evolve when oxygen levels in the atmosphere rose to close to modern levels. But now studies of a small sea sponge fished out of a Danish fjord shows that complex life does not need high levels of oxygen in order to live and grow.The origin of complex life is one of science's greatest mysteries. How could the first small primitive cells evolve into the diversity of advanced life forms that exists on Earth today? The explanation in all textbooks is: Oxygen. Complex life evolved because the atmospheric levels of oxygen began to rise app. 630 -- 635 million years ago.However new studies of a common sea sponge from Kerteminde Fjord in Denmark shows that this explanation needs to be reconsidered. The sponge studies show that animals can live and grow even with very limited oxygen supplies.In fact animals can live and grow when the atmosphere contains only 0.5 per cent of the oxygen levels in today's atmosphere."Our studies suggest that the origin of animals was not prevented by low oxygen levels," says Daniel Mills, PhD at the Nordic Center for Earth Evolution at the University of Southern Denmark.Together with Lewis M. Ward from the California Institute of Technology he is the lead author of a research paper about the work in the journal PNAS.A little over half a billion years ago, the first forms of complex life -- animals -- evolved on Earth. Billions of years before that life had only consisted of simple single-celled life forms. The emergence of animals coincided with a significant rise in atmospheric oxygen, and therefore it seemed obvious to link the two events and conclude that the increased oxygen levels had led to the evolution of animals."But nobody has ever tested how much oxygen animals need -- at least not to my knowledge. Therefore we decided to find out," says Daniel Mills.The living animals that most closely resemble the first animals on Earth are sea sponges. The species Halichondria panicea lives only a few meters from the University of Southern Denmark's Marine Biological Research Centre in Kerteminde, and it was here that Daniel Mills fished out individuals for his research."When we placed the sponges in our lab, they continued to breathe and grow even when the oxygen levels reached 0.5 per cent of present day atmospheric levels," says Daniel Mills.This is lower than the oxygen levels we thought were necessary for animal life.The big question now is: If low oxygen levels did not prevent animals from evolving -- then what did? Why did life consist of only primitive single-celled bacteria and amoebae for billions of years before everything suddenly exploded and complex life arose?"There must have been other ecological and evolutionary mechanisms at play. Maybe life remained microbial for so long because it took a while to develop the biological machinery required to construct an animal. Perhaps the ancient Earth lacked animals because complex, many-celled bodies are simply hard to evolve," says Daniel Mills.His colleagues from the Nordic Center for Earth Evolution have previously shown that oxygen levels have actually risen dramatically at least one time before complex life evolved. Although plenty of oxygen thus became available it did not lead to the development of complex life.据外媒2⽉17⽇报道，科学界⼀直认定，地球上的复杂⽣物是在⼤⽓中的氧⽓含量达到现在的标准时才逐渐进化出来的。
关于水杨酸研究的英文文献
关于水杨酸研究的英文文献Abstract:Salicylic acid (SA), primarily derived from willow bark, has been extensively studied due to its diverse therapeutic effects on human health and its potential applications in various fields. This review aims to explore the recent advancements in salicylic acid research and summarize the key findings and applications of SA in medicine, skincare, agriculture, and other domains. The paper also addresses the mechanisms of action and potential side effects associated with salicylic acid usage.1. IntroductionSalicylic acid, a monohydroxybenzoic acid, has been used in various forms for centuries due to its analgesic, antipyretic, anti-inflammatory, and antiplatelet properties. This section briefly discusses the history, chemical properties, sources, and forms of salicylic acid.2. Pharmacological Properties of Salicylic AcidThis section focuses on the pharmacological properties of salicylic acid, including its analgesic, anti-inflammatory,anti-platelet, and antipyretic effects. Various studies are discussed to understand the underlying mechanisms of these properties.3. Medical ApplicationsSalicylic acid has been widely used in the medical field for treating various conditions, such as pain management, acne, psoriasis, and warts. This section reviews the clinical efficacy of salicylic acid in these applications based on available studies, and also highlights the challenges and future prospects of salicylic acid-based therapeutics.4. Skincare and Cosmetic ApplicationsThe use of salicylic acid in skincare and cosmetic products has gained significant popularity due to its effectiveness in treating acne, reducing hyperpigmentation, and improving the overall appearance of the skin. This section discusses the formulation and efficacy of salicylic acid formulations in skincare products, including cleansers, toners, creams, and masks.5. Agriculture ApplicationsSalicylic acid has emerged as a potential plant growth regulator, providing enhanced resistance against various biotic and abiotic stresses. This section explores the role ofsalicylic acid in agriculture, including its effects on plant growth, disease resistance, and post-harvest preservation.6. Mechanisms of ActionUnderstanding the mechanisms of action is essential for harnessing the potential of salicylic acid. This sectiondiscusses the cellular and molecular pathways involved in the pharmacological effects of salicylic acid, such as its regulation of cyclooxygenase (COX) enzymes, NF-κB signaling, and reactive oxygen species (ROS) generation.7. Side Effects and PrecautionsWhile salicylic acid has diverse therapeutic effects, it is essential to understand its potential side effects and precautions. This section examines the adverse reactions associated with salicylic acid usage, such as gastrointestinal irritation, bleeding, and allergic reactions. Additionally, it provides guidelines to ensure safe and effective use ofsalicylic acid.8. ConclusionKeywords: Salicylic acid, medical applications, skincare, agriculture, mechanisms of action, side effects.。
单唾液酸四己糖神经节苷脂钠联合脑蛋白水解物治疗帕金森病的疗效观察
单唾液酸四己糖神经节苷脂钠联合脑蛋白水解物治疗帕金森病的疗效观察发表时间：2019-07-24T15:59:14.347Z 来源：《中国结合医学杂志》2019年5期作者：朱少平[导读] 针对临床收治的帕金森病患者给予单唾液酸四己糖神经节苷脂钠联合脑蛋白水解物治疗，可有效增强临床总有效率，缩短症状改善时间，安全性较高，具有临床研究意义。
娄底市双峰县人民医院湖南娄底 417700【摘要】目的：观察单唾液酸四己糖神经节苷脂钠联合脑蛋白水解物治疗帕金森病的临床疗效。
方法：选取临床收治的帕金森病患者56例，均为2017年1月至2018年12月期间收治，就单纯给予单唾液酸四己糖神经节苷脂钠治疗（对照组，n=28）与给予单唾液酸四己糖神经节苷脂钠联合脑蛋白水解物治疗（观察组，n=28）临床疗效、治疗前后UPDRS评分及不良反应展开对比。
结果：在临床疗效、治疗后UPDRS评分的对比上，观察组均优于对照组（P<0.05）；治疗后两组均发生1例不良反应，组间比较无差异（P>0.05）。
结论：针对临床收治的帕金森病患者给予单唾液酸四己糖神经节苷脂钠联合脑蛋白水解物治疗，可有效增强临床总有效率，缩短症状改善时间，安全性较高，具有临床研究意义。
【关键词】帕金森病；单唾液酸四己糖神经节苷脂钠；脑蛋白水解物；疗效【Abstract】Objective：To observe the clinical efficacy of monosialotetrahexosyl ganglioside combined with brain protein hydrolysate in the treatment of Parkinson's disease. METHODS：A total of 56 patients with clinically treated Parkinson's disease were enrolled. All patients were treated with monosialotetrahexosylganglioside sodium from January 2017 to December 2018（control group，n=28）. The clinical efficacy，UPDRS score and adverse reactions before and after treatment were compared with the administration of monosialotetrahexosylganglioside sodium combined with brain protein hydrolysate（observation group，n=28）. RESULTS：In the comparison of clinical efficacy and post-treatment UPDRS scores，the observation group was superior to the control group（P<0.05）. One patient had adverse reactions after treatment，and there was no difference between the two groups（P>0.05）. Conclusion：The treatment of patients with Parkinson's disease treated with monosialotetrahexosylganglioside combined with brain protein hydrolysate can effectively enhance the total clinical effective rate，shorten the symptom improvement time，and have high safety. significance.【Key words】 Parkinson's disease；monosialotetrahexosylganglioside sodium；brain protein hydrolysate；therapeutic effect帕金森病属于神经系统疾病，多发生在中老年人群体，随着我国人口逐渐趋向老龄化，工作生活压力增大，帕金森病逐年上升，成为神经内科较为常见的疾病[1]。
嗅成鞘细胞海马内注射对阿尔茨海默病模型大鼠的作用
阿 尔 茨 海 默 病 （ ｚ ｅ ｒＳｄｓａｅ ＡＤ） Ａ１ｈ ｉ ｉｅｓ ， ｍｅ 是
一
Hale Waihona Puke 探讨 Ｏ Ｃ 移 植 对 阿 尔 茨海 默 病 模 型 大 鼠 的治 疗 Ｅ ｓ
效果 。
组 以学 习记忆 减 退 和认 知 障 碍 为 主 要 特 点 的 临
床 综合 征 ， 发病 机 制与 神经 元 中的线 粒体 结 构受 其 损 和能 量代 谢 障碍关 系密 切 ； 细胞 色素 氧 化酶 而 （ｙｏ ｈ ｏ ｘｄ ｓ ， ＯＸ） ｃｔｃ ｒｍｅｏ ｉａｅ Ｃ 又是 线 粒 体 呼 吸 链 的
４ Ｏ只 随 机 分 为 ４组 ： 康 对 照 组 、 健 ＡＤ 模 型 组 、 Ｃ 移 植 ＯＥ ｓ 组、 Ｓ ＭＣ Ｆ注 射 组 ， 组 １ 每 ０只 １２ 建 立 大 鼠 Ａ ． Ｄ模 型 将 Ａ ． 于无 菌 生 理 盐 水 （ ￣ ／ ， 人 ３ ℃ 温 Ｂ ． 溶 ５／ ｍＩ） 放 ｇ ７
郧 阳 医学 院 （ ４ ０ ０ ４２０） 姚 淞 元 姚 伟 史 丹 青 王金 勇 王 军 姚 柏 春
【 摘 要 】 目 的 探 讨 嗅 成 鞘 细 胞 （ Ｅ ｓ海 马 内注 射 对 阿 尔茨 海 默 病模 型 大 鼠 的作 用 。 方 法 Ｓ 大 鼠双 Ｏ Ｃ） Ｄ 侧 海 马注 射 Ａａ ， 立 ＡＤ 大 鼠模 型 。 实 验 动 物 分 为 ４组 ： 康 对 照 组 、 ｌ 。 建 健 ＡＤ模 型 组 、 Ｃ 移 植 组 、 工 脑 脊 液 ＯＥ ｓ 人 注射组 ， 组 ｌ 每 Ｏ只 。 体外 原 代 培 养 嗅 成 鞘 细 胞 并 将 其 移 植 至 Ａ 大 鼠海 马 内 。运 用 行 为 学 测 试 、 织 化 学 、 位 Ｄ 组 原 杂 交 ， 合 图 像 分 析 以 及 电镜 等 技 术 ， 察 、 结 观 比较 各 组 大 鼠学 习 记 忆 能 力 、 马 Ｃ 区线 粒 体 细 胞 色 素 氧 化 酶 海 Ａ
immunohistochemistry labelling 免疫组化
Fluorescent labels
Advantages
Size Specificity Sensitivity
Disadvantages
Hardware Limited selection Background
IgG
IgG
Confocal
Application in renal diseases
Very high sensitivity Used in research more than routine studies. It is longer and more expensive.
• In order to visualize the enzymes labelling the antibodies with light microscope, enzyme – substrate reactions, which convert colorless chromogens into visible colored endproducts, is used.: • Peroxidase- hydrogen peroxide- diaminobenzidine (DAB): BROWN
IgG
The basic principle of immunofluorescence
To
use a fluorescent compound (usually fluorescein) to detect the binding of antigen and antibody
The Ab is labelled with the fluorescent compound Under a fluorescence microscope, fluorescein