Clearing and embedding in polyester resin for demonstrating the nerve distribution pattern of skeletal muscles

The preservation of many stained gross specimens in solution creates some difficulties. It is convenient and effective to preserve material in polyester resin instead of glycerol. The aim of this study was to determine the usefulness of clearing and embedding using polyester resin. The samples consisted of the nerve distribution patterns of skeletal muscles stained using Sihler's method. The muscles were cleared more successfully and the intramuscular nerve distributions were demonstrated better in polyester than in glycerol. The method presented here eliminates not only the storage and handling problems of specimens, but also problems such as pale stains and the molding of preparations. Furthermore, it is more convenient to examine and to photograph specimens cleared and embedded in polyester than those stored in glycerol.     

Staining and Mounting Helminths

The following procedure is recommended: Fix ces-todes and trematodes (while held flat between glass slides) 0.5–2.0 hr. in the following mixture: formalin, 15; acetic acid (gl.), 5; glycerol, 10; 95% ethyl alcohol, 24; distilled H₂O, 46; all proportions by volume. After freeing them from the slides, wash thoroughly in running water and stain immediately thereafter. Stock staining solution: ferric ammonium alum (violet cryst.), 2 g.; distilled H₂O (cold) 100 ml.; after solution, add 2 ml. concentrated H₂SO₄, bring to a boil; add 1 g. coelestin blue B (Nat. Aniline), boil 3–5 min.; cool and add 10 ml. absolute methyl alcohol and 10 ml. glycerol. Dilute 1 vol. with 3 vol. distilled H20 for use. Stain 5–30 min., depending on size of specimens. Wash with 2 changes 0.5 hr. each of distilled H₂O, then 50% isopropyl alcohol 12–16 hr., 50% isopropyl alcohol 2 hr., followed by graded isopropyl alcohol for dehydration. Ether: ethyl alcohol (equal parts), 1 hr., is followed by embedding in celloidin in a sheet just thick enough to cover the specimens. Trim embedded specimens and dehydrate with isopropyl alcohol, 80%, 90% and absolute. Clear in beechwood creosote. Mount in balsam with cover glasses that overlap the edges of the celloidin 1–2 mm. While drying at 37°C, refill edges of mount with fresh balsam as needed. When dry, remove excess balsam and ring the edges with ordinary gloss enamel paint.     

穴位埋线联合补肾调经汤对多囊卵巢综合征患者性激素水平及代谢指标的影响

摘要 目的：探讨穴位埋线联合补肾调经汤对多囊卵巢综合征（PCOS）患者性激素水平及糖脂代谢指标的影响。方法：选择2018年10月至2019年9月广州中医药大学附属宝安区中医院妇科门诊收治的116例PCOS患者,按随机数字表法将患者分为两组,各58例。对照组采用炔雌醇环丙孕酮片治疗3个月,观察组给予补肾调经汤和穴位埋线治疗3个月。观察两组疗效、治疗前后体质量指数（BMI）、性激素水平和糖脂代谢指标的差异。结果：观察组治疗总有效率高于对照组（P＜0.05）。两组治疗后BMI、睾酮（T）、促黄体生成素（LH）均下降,且观察组低于对照组（P＜0.05）,两组治疗后促卵泡激素（FSH）比较无统计学差异（P＞0.05）。两组治疗后总胆固醇（TC）、甘油三酯（TG）、低密度脂蛋白胆固醇（LDL-C）、空腹血糖（FPG）、空腹胰岛素（FINS）、胰岛素抵抗指数（HOMA-IR）均有所下降,且观察组低于对照组,高密度脂蛋白胆固醇（HDL-C）有所升高,且观察组高于对照组（P＜0.05）。结论：穴位埋线联合补肾调经汤治疗PCOS的疗效满意,能够控制体重,可改善PCOS患者性激素水平和糖脂代谢紊乱。     

Preparing sagittae for examination of daily growth increments of young‐of‐the‐year fishes: a modification of the embed method

A modification (termed the slide‐glass‐embed‐method, SGEM) of the embed method for preparing fish sagittae is described. The SGEM is based on a very simple principle: a dome of mixed resin containing the embedded sagittae loses hardness after being heated and can be easily cut with dissecting scissors.     

基于核局部线性嵌入的基因表达谱数据分类

针对局部线性嵌入算法（LocalLinearEmbedding,LLE）利用试凑法寻找近邻数耗时的缺陷性,提出一种增强的核局部线性嵌入算法（EnhancedKernelLocalLinearEmbedding,EKLLE）自动为样本分配邻域;该算法以高斯核函数为核心改进标准LLE距离度量准则,结合样本的类别信息,无需人工干预自动为样本设置不同的近邻数,克服了试凑法获得最优结果时需要大量时间;最后在各样本近邻数不相同的情况下对数据进行维数简约及待测样本分类。EKLLE算法有效地将高维基因表达谱数据映射到低维本质空间中,解决了传统LLE算法不能很好地处理合噪声或者稀疏数据的缺点。通过对比其他肿瘤样本分类实验,验证本文方法的实时性和精确性。     

采用常规电泳设备建立CLARITY透明脑技术

目的:CLARITY是一种全新的组织形态学研究技术,在脑结构与功能研究中具有重要用途。探讨能否采用常规电泳设备开展该项技术。方法:从ICR雌性小鼠取得新鲜脑组织,依次进行多聚甲醛组织固定、聚丙烯酰胺水凝胶包埋、SDS电泳洗涤,通过拍照记录结果。结果:多聚甲醛固定和水凝胶包埋后的脑组织呈乳白色,柔软富有弹性;电泳洗涤后的脑组织从周边开始逐渐透明化,至洗涤第10 d时呈现均匀的半透明状态,可用于三维神经网络原位分析。结论:研究结果为采用常规电泳设备开展CLARITY技术提供了基础数据。     

温度对兰州百合鳞片埋培繁殖的影响

为了筛选出兰州百合鳞片埋培繁殖的最适温度和鳞片层次,解决兰州百合种源不足、繁育周期长的问题,该文以兰州百合鳞片为材料,采用温度(20、25、30℃)和鳞片层次(外层、中层、内层)二因素完全随机区组设计,研究了二因素对兰州百合鳞片埋培繁殖效果的影响。通过对鳞片疑似发病率、分化率及小鳞茎分化数进行统计与分析,结果发现不同温度处理及各鳞片层次对鳞片疑似发病率、分化率及小鳞茎分化数的影响存在显著或极显著差异。结果表明:(1)温度越高,鳞片的疑似发病率越低,在埋培2周时,20℃处理下疑似发病率最高(38.67%),30℃处理下最低(10%);各层次鳞片的疑似发病率由高到低依次为外层中层内层。(2)在25、30℃处理下,小鳞茎分化率最高,埋培结束(6周)时分别为91.33%、90.89%;中层及内层鳞片小鳞茎分化率极显著高于外层鳞片。(3) 30℃处理下鳞片形成小鳞茎数最多,在埋培6周时达到每片2.00粒;同时中层及内层鳞片小鳞茎分化数显著高于外层鳞片。综上结果表明,兰州百合鳞片埋培繁殖以选用中层(3~4层)、内层(5~7层)鳞片在25~30℃条件下繁殖效果最好。     

Three-layer ONIOM studies of the dark state of rhodopsin: the protonation state of Glu181

A computational three-layer ONIOM(QM-high:QM-low:MM) hybrid scheme has been applied to analyze the protonation state of the Glu181 amino acid residue in rhodopsin, which is vital to determining the rhodopsin photoactivation mechanism. Due to conflicting evidence from previous studies, it has yet to be conclusively resolved. In this study, we fully optimize dark-state rhodopsin model structures differing only at the 181-residue site—protonated and unprotonated Glu181—and calculate several experimentally observable properties. Comparison of calculated structures, excitation energies, and NMR chemical shifts for the two models with values from the literature allows a reevaluation of previously reported conclusions. A key finding is that the S₁ → S₂ energy level splitting, previously used as evidence for a neutral Glu181, is found to be almost identical for the two protonation states. We highlight a need for caution when interpreting experimental data. Small differences in the properties of the two model structures are also identified, which may be useful targets for future high-resolution experimental approaches.     

胡萝卜人工种子包衣材料的筛选

以胡萝卜SK4-316种子苗下胚轴为外植体诱导培养的体细胞胚,进行人工种子的制作。结果表明,海藻酸钠包埋体系制作的人工种子萌发率较稳定,PEG体系的萌发率高于前者,但萌发后成苗率较低。以MS液体培养基作包埋培养基,4%海藻酸钠、2% CaCl₂·2H₂O、0.1 mg/L GA₃、0.2%活性炭组成的海藻酸钠复合包衣剂制作的人工种子,在1/16MS培养基、24℃下培养萌发率最高,可达97.02%。