不同固定液和染色方法显示山羊子宫内肥大细胞效果的比较

目的探讨用不同固定液和染色方法对显示处于间情期山羊子宫肥大细胞的影响。方法用四种不同的固定方法,应用改良甲苯胺蓝（MTB）染色法和阿尔辛蓝-番红花红（AB-S）染色法显示处于间情期山羊子宫肥大细胞。结果山羊子宫组织采用Carnoy氏液固定,MTB和AB-S染色对所有的肥大细胞均可获得良好的染色反应,但10％中性福尔马林,4％多聚甲醛,Bouin氏液固定的组织仅有少量肥大细胞着染。结论MTB和AB-S染色法均是山羊子宫肥大细胞良好的染色方法。     

恶性肿瘤中抗酸菌L型检出意义的研究

本文用改良抗酸染色法和卡介苗抗体免疫组化染色法对４２０例恶性肿瘤（乳腺癌１１２例肠腺癌１００例、恶性淋巴病５４例、子宫颈癌７０例、绒毛膜癌８４例）进行了研究。结果发现４２０例恶性肿瘤中４７例（１１．２％）,１Ｋ抗酸染色阳性和４９例（１１．６％）免疫组化染色阳性,其中以恶性淋巴瘤抗酸菌Ｌ型检出率最高（４４．４％）绒毛膜癌最低（２．４％）,抗酸菌Ｌ型主要（８５％）分布于癌巢或肿瘤实质内该区５１．１％癌细胞呈增生活跃状态,３４％癌细胞呈空泡样变性,颇似病毒感染的凹空细胞,２．１％的瘤细胞发生坏死。结合文献复习,提示抗酸菌Ｌ型感染可能与肿瘤发生有关。     

改良PAS染色法在急性肝损伤中的应用

目的：观察改良肝脏糖原PAS染色法,并观察肝脏糖原染色在急性肝损伤中的应用。方法：复制CCl4急性肝损伤模型,首次100％CCl43 mL／kg皮下注射,此后50％CCl4橄榄油溶液2 mL／kg每周2次共4次皮下注射,诱导大鼠急性肝损伤模型。计算大鼠肝体比;HE染色观察肝组织炎症病理;试剂盒检测血清丙氨酸氨基转移酶（ALT）、天门冬氨酸氨基转移酶（AST）、总胆红素（TBil）、白蛋白（Alb）。肝脏常规PAS染色与改良PAS染色观察肝糖原染色。结果：与正常组相比,模型组ALT、AST活性与TBil含量明显升高（P＜0．05）,Alb含量明显降低（P＜0．05）;HE染色示,模型组肝小叶结构排列紊乱,肝细胞脂肪变、气球样变明显。常规PAS染色,正常组肝组织PAS染色阳性占肝脏面积为32．38％±5．50％;与正常组相比,模型组肝组织PAS阳性染色明显减少（P＜0．01）,占肝脏面积为8．60％±3．34％。改良PAS染色提示,正常组肝脏可见大量PAS阳性染色,占肝脏面积为75．50％±9．02％;与正常组相比,模型组肝组织PAS阳性染色明显减少（P＜0．01）,占肝脏面积为17．61％±3．53％。在空白对照组与模型肝组织中,肝糖原改良PAS染色阳性率明显高于常规PAS染色法（P＜0．01）。改良PAS染色肝糖原阳性染色面积更真实反映急性肝损伤程度。结论：改良肝脏糖原PAS染色法有助于急性肝损伤程度评估。     

植物细胞壁组织化学定位染色方法和技术的比较研究

利用光学和荧光显微镜比较研究几种植物细胞壁组织化学定位染色方法和技术,结果表明：（1）硫酸消化法和硫酸氢黄连素-苯胺兰对染法研究凯氏带,对取材时间和部位要求高,建议两种方法配合使用,可相互印证是否具凯氏带;（2）苏丹7B染色法,蓝色激发光下不染色和硫酸氢黄连素-苯胺兰对染研究细胞壁栓质层3种方法中,不染色蓝色激发光下结果比苏丹7B染色法敏感显色,但苏丹7B染色法在普通光学显微镜下观察较为便捷;（3）木质化细胞壁染色方法中硫酸氢黄连素-苯胺兰对染法比间苯三酚-盐酸染色法易显色观察;（4）甲苯胺兰快速染色细胞壁取代常规苏丹Ⅲ/Ⅳ法,细胞边界和层次更清楚。     

改良革兰染色法在显示石蜡切片真菌中的应用

目的建立真菌的病理切片改良革兰染色法。方法选取确诊真菌感染的组织蜡块,切若干空白片,通过苏木素-伊红染色（HE）、高碘酸-无色品红染色（PAS）、六胺银染色（GMS）、改良革兰染色后,比较改良革兰染色法的染色效果。结果改良革兰染色法的染色效果好,该法具有易操作、染色效果稳定等优点。结论改良革兰染色法能够清晰地显示出组织切片中的真菌,并且染色效果稳定,在检测组织中的真菌时可发挥重要的辅助诊断价值,具有广泛的应用前景。     

观察"胞间连丝"染色方法的改进

在大部分的高校观察“胞间连丝”都是观察现成的永久装片 ,而在一般的专科院校及中学 ,由于条件的限制 ,缺乏现成的装片 ,大都选用柿核的胚乳作实验材料 ,进行徒手切片和染色来观察。笔者对染色的方法作了一些改进 ,介绍如下 :1　水装片的制作取新鲜的柿核 ,除去种皮 ,取一窄条用刀片作连续的徒手切片 ,取较薄的切片制成水装片 ,用显微镜检查。2　染色方法和观察结果1 )装片用质量分数 0 .1 %龙胆紫溶液染色 5～ 1 0min ,水洗、吸干 ,再用质量分数 0 .1 %酸性品红复染 5～1 0min ,水洗、镜检 ,可见柿核胚乳细胞壁染成了浅红色(或带浅紫色 ) …     

核酸染料Genefinder使用方法的比较

目的:使用核酸染料Genefinder检测琼脂糖凝胶中的核酸,通过比较预染样品法、胶内染色法和后染法三种染色方法的染色效果,了解该染料的染色特性,以期找到性能稳定,染色效果好的染色方法.方法:在琼脂糖凝胶电泳中,以不同的染色方法使用核酸染料Genefinder进行染色,对染色结果进行比较分析.结果:使用电泳后染色方法染色效果较好,胶内染色法次之,预染样品法效果最差.结论:核酸染料Genefinder会干扰DNA的迁移效率,因此,使用Genefinder进行电泳后染色是一种较好的染色方法.     

消化道组织块黏液细胞的组织化学染色

目的：探索中华蟾蜍、黑斑蛙消化道组织块黏液细胞的组织化学染色。方法：利用阿辛蓝-过碘酸Schiff（AB-PAS）反应,对消化道组织块黏液细胞进行染色和观察。结果：对消化道壁较薄的组织块（食管、小肠、大肠）,采用3％乙酸3min、1％阿尔新蓝30min、3％乙酸3min、3％过碘酸氧化10min、Schiff反应20min染色,即可达到良好染色效果;对消化道壁较厚的组织块（胃）,则应采用3％乙酸9min、1％阿尔新蓝70min、3％乙酸9min、3％过碘酸氧化30min、Schiff反应60min染色,将能达到良好染色效果。结论：经过染色可将黏液细胞分为四个类型：Ⅰ型红色,PAS染色阳性,AB染色阴性;Ⅱ型蓝色,PAS染色阴性,AB染色阳性;Ⅲ型紫红色,PAS染色强阳性,AB染色弱阳性;Ⅳ型蓝紫色,PAS染色弱阳性,AB染色强阳性。     

石榴皮染料对毛织物的染色性能研究

本文研究了资源丰富的石榴皮染料溶液的耐热水稳定性,分析了染浴pH值、染色温度对石榴皮染料上染毛织物的影响,并测试了所染毛织物的色牢度及抗紫外性能。研究结果表明,石榴皮染料溶液耐热水稳定性好。石榴皮染料可采用直接染、铝、铁离子预媒染和后媒染多种染色方法对毛织物染色,不同染色方法可得到不同颜色的毛织物。直接染色法最佳染浴pH值为3,铁后媒染色法最佳染浴pH值为7,各种染色方法染色温度应为100℃。石榴皮染料染色毛织物具有良好的皂洗牢度、摩擦牢度和升华牢度,并具有很强的抗紫外性能。石榴皮染料可作为毛织物的一种功能型天然染料。     

胃腺癌血管生成拟态和HIF—1α的表达及意义

目的探讨在人胃腺癌组织中是否存在血管生成拟态（Vasculogenic mimicry,VM）及VM与HIF-1α表达的关系,以及HIF—1α表达的临床病理意义。方法收集临床病例资料完整的胃腺癌121例,利用CD34和PAS双重染色观察是否存在VM,然后对VM阳性组和阴性组进行HIF-1α染色,分析VM与HIF-1α表达的关系及HIF—1α表达与患者临床病理指标的关系。结果121例胃腺癌组织中有44例（36．36％）存在VM。VM阳性组和VM阴性组HIF-1α蛋白表达阳性率分别为77．27％（34／44）和54．55％（42／77）,两组间的差异有统计学意义（P〈0．05）;有、无淋巴结转移的胃腺癌组织HIF-1α表达阳性率分别为76．74％（66／86）和28．57％（10／35）,两组间的差异有显著性（P〈0．01）;中、低分化胃腺癌组织中HIF-1α表达阳性率分别为50．85％（30／59）、74．19％（46／62）,两组间的差异有显著性（P〈0．01）。结论胃腺癌组织中存在VM,HIF-1α的表达可能促进VM的形成。VM与HIF-1α的表达可能是胃腺癌浸润、转移的重要生物学标志。     

小球藻Rubisco在叶绿体中的免疫金标定位

应用免疫技术对Rubisco在中国小球藻(Chlorellaspp.640909)叶绿体中进行了分子定位及Native-PAGE电泳、SDS-PAGE电泳及其Westen印迹分析,并对小球藻淀粉核(Pyrenoid)超微结构进行了观察.结果显示Native-PAGE电泳图谱主要为一条主带,Westen印迹反应证明该条带即为Rubisco酶,SDS-PAGE电泳及其Western印迹图谱显示Rubisco大亚基分子量大约为55kD.中国小球藻淀粉核为椭圆形,被淀粉鞘所包围,中央有一条由2个类囊体组成的纵向通道,并在蛋白核内段处稍膨胀.淀粉核与叶绿体基质存在多处联系.免疫分子定位显示Rubisco大亚基和全酶分子主要分布于叶绿体的淀粉核上,且Rubisco在淀粉鞘部位也有少量分布,极少部分分布在叶绿体基质中,表明叶绿体淀粉核与光合作用关系密切.Rubisco聚集于淀粉核可能有利于藻类对CO2固定.     

Immunogold localization of ribulose-1,5-bisphosphate carborylsae/oxygenase in chloroplasts of Chlorella]

Ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) is a first key enzyme in the Calvin Circle of plant cell photosynthesis. This paper mainly studied gold immunolocalization of Rubisco of Chlorella spp. 640909, and the Native-PAGE and, SDS-PAGE and Western bloting analysis, as well as the observation to pyrenoid ultra structure. The Native-PAGE result showed a main band, evidenced as the Rubisco band by the Western blot with the antibody against the Rubisco from C. prototothecoides, The special immunoacton of Rubisco from Chlorella spp. 640909 and the antibody to large subunit of Rubisco from C. prothecoides showed the large subunit proteins of Rubisco in the two species of Chlorella shared the high homology. The SDS-PAGE and Western blotting maps showed the molecule weight of the large subunit of Rubisco of Chlorella spp. 640909 was about 55 KD. The shape of pyrenoid ultra structure of the electronic microscope was oblong, and was embedded in starch sheath, with 2 swelling thylakoids through out a center portrait channel of the pyrenoid. There were some connections between pyrenoid and the chloroplast stroma. The distribution of the large subunits and the whole Rubisco in the chloroplast of Chrolella spp. 640909 was studied by immunoelectron microscopy by embedded sections with antibody to large subunit and whole enzyme followed by second antibody, goad anti-rabbit immunoglobulin G conjugated to 10 nm gold particles(Sigma production). The result showed the antibodies against large subunit and whole enzyme heavily labeled the pyrenoid, as well as starch sheath region, whereas the thylakoid region of the plastid was lightly labeled. And the whole Rubisco antibody labeled the pyrenoid surface more heavily than the large subunit antibody did. It is demonstrated the pyrenoid and starch sheath have the photosynthesis function. Rubisco concentrating in pyrenoid and starch sheath is valuable to fix CO2 for photosynthesis in algae.     

The induction of the CO2-concentrating mechanism is correlated with the formation of the starch sheath around the pyrenoid of Chlamydomonas reinhardtii

The pyrenoid is a prominent proteinaceous structure found in the stroma of the chloroplast in unicellular eukaryotic algae, most multicellular algae, and some hornworts. The pyrenoid contains the enzyme ribulose-1,5-bisphosphate carboxylase/oxygenase and is sometimes surrounded by a carbohydrate sheath. We have observed in the unicellular green alga Chlamydomonas reinhardtii Dangeard that the pyrenoid starch sheath is formed rapidly in response to a decrease in the CO₂ concentration in the environment. This formation of the starch sheath occurs coincidentally with the induction of the CO₂-concentrating mechanism. Pyrenoid starch-sheath formation is partly inhibited by the presence of acetate in the growth medium under light and low-CO₂ conditions. These growth conditions also partly inhibit the induction of the CO₂-concentrating mechanism. When cells are grown with acetate in the dark, the CO₂-concentrating mechanism is not induced and the pyrenoid starch sheath is not formed even though there is a large accumulation of starch in the chloroplast stroma. These observations indicate that pyrenoid starch-sheath formation correlates with induction of the CO₂-concentrating mechanism under low-CO₂ conditions. We suggest that this ultrastructural reorganization under lowCO₂ conditions plays a role in the CO₂-concentrating mechanism C. reinhardtii as well as in other eukaryotic algae.     

几种藻类蛋白核的超微结构研究

应用电镜及免疫电镜技术对莱茵藻、小球藻、条浒苔和紫菜等藻类的叶绿体蛋白核的超微结构及主要组成成分进行了观察和研究。结果显示：不同藻类的蛋白核结构不同,显示了藻类蛋白核的多样性。蛋白核为球形或椭圆形,由蛋白质组成。莱茵藻、小球藻和条浒苔的蛋白核外围被淀粉鞘所包围,而紫菜的蛋白核外围无淀粉鞘而直接被叶绿体的类囊体所包围。淀粉鞘由淀粉组成,淀粉鞘的厚薄与藻体藻龄及增养状态有关系。在蛋白核中央,一般都具有由类囊体形成的孔道,使蛋白核与外界联系,小球藻和条浒苔蛋白核具有1条纵向孔道,而莱茵藻和紫菜为多条孔道。金相免疫技术检测结果显示Rubisco和Rubisco活化酶均在蛋白核及淀粉鞘区域中定位,表明蛋白核具有光合作用功能．     

Immunocytochemical Localization of Phosphoribulokinase in Microalgae

Employing immunogold electron microscopy, the subcellular location of the Calvin cycle enzyme phosphoribulokinase (PRK) was determined for two diverse species of microalgae. In both the red alga Porphyridium cruentum and the green alga Chlamydomonas reinhardtii, PRK was distributed throughout the thylakoid-containing chloroplast stroma. In contrast, the next enzyme in the pathway, ribulose 1,5-bisphosphate carboxylase/oxygenase, was predominantly pyrenoid-localized in both species. In Porphyridium, the chloroplast stroma abuts the pyrenoid but in Chlamydomonas and other green algae, the pyrenoid appears encased in a starch sheath. Unique inclusions found in the pyrenoid of Chlamydomonas were immunolabelled by anti-PRK and thus identified as regions of chloroplast stroma. It is postulated that such PRK-containing stromal inclusions in the pyrenoids of Chlamydomonas and perhaps other green algae provide a means for exchange of Calvin cycle metabolites between pyrenoid and stroma.     

The induction of the CO2 concentrating mechanism in a starch-less mutant of Chlamydomonas reinhardtii

In Chlamydomonas reinhardtii the formation of a starch sheath surrounding the pyrenoid is observed when cells grown under high CO₂ (5% CO₂ in air) are transferred to low CO₂ (0.03%) conditions. Formation of the starch sheath occurs coincidentally with induction of the CO₂ concentrating mechanism and with de novo synthesis of 5 polypeptides with molecular masses of 21, 36, 37, 42–44 kDa. We studied the effect of CO₂ concentrations on photosynthesis, ultrastructure and protein synthesis in Chlamydomonas reinhardtii cw-15 (wild phenotype for photosynthesis) and in the starch-less mutant BAFJ -6, with the aim to clarify the role of the pyrenoid starch sheath in the operation of the CO₂ concentrating mechanism and whether these low CO₂-inducible polypeptides are involved in the formation of starch sheath. When wild type and starch-less mutant cells were transferred from high to low CO₂, the CO₂ requirement for half-maximal rates of photosynthesis decreased from 40 μM to 2 μM CO₂. ³⁵SO₄^2- labeling analyses showed that the starch-less mutant induced the 5 low CO₂-inducible polypeptides. These observations suggest that the starch-less mutant was able to induce a fully active CO₂ concentrating mechanism. Since the starch-less mutant did not form a pyrenoid starch sheath, we suggest that the starch sheath is not involved in the operation of the CO₂ concentrating mechanism and that none of these 5 low CO₂-inducible proteins is involved in the formation of the starch sheath in Chlamydomonas .     

Studies on Enzymatic Liquefaction and Saccharification of Starch

It was confirmed that turbid substances which remained in saccharified liquid after digestion of starch slurry with bacterial alpha-amylase and glucoamylase were insoluble starch particles and that these materials were formed during heat treatment of the slurry in liquefaction process.

In iodine staining reaction, the suspension of these materials showed only a slightly dirty blue color; even after boiling of this suspension for 30 min at 135°C, this staining reaction was not complete and the maximum color intensity could be obtained after treatment with 1 n caustic soda solution.

It is assumed from the iodine color reaction, that the main component of these materials was glucose polymer of straight chain, like that of amylose, and that this would be strongly associated with each other to form the compact molecule.

The method for determination of these insoluble starch particles in the starch slurry was established and by this method the contents of these materials were determined in some types of starches.     

尼罗红荧光法快速检测培养过程中湛江等鞭金藻的中性脂

研究一种快速准确测定微藻中中性脂的方法。湛江等鞭金藻是一种中性脂含量高且具有开发潜力的能源微藻。以湛江等鞭金藻为实验对象,首先优化尼罗红染色的条件。当二甲基亚砜体积分数为2.0%、尼罗红质量浓度为1.00μg/m L、细胞密度为1.0×106个/m L、激发波长为480 nm、检测波长为580 nm时,优化的染色时间为10min。其次测定了背景荧光对检测的影响。结果表明,在不同细胞状态下,背景荧光强度大约是微藻内荧光强度的20%左右,可以忽略。最后比较了尼罗红荧光法和重量法。结果表明,荧光强度与中性脂含量的相关系数R2=0.946 8,虽然两者相关性并不十分高,但作为一种快速测定微藻中中性脂的方法,尼罗红荧光法依然是研究微藻培养过程中中性脂含量变化的有效方法。     

深山含笑叶片总酚超声波提取工艺的优化

采用单因素实验方法研究了超声波辅助提取过程中乙醇浓度、料液比、提取温度和提取时间对深山含笑（Michelia maudiae Dunn）叶片总酚提取率的影响,并采用正交实验方法确定了最佳提取工艺条件。结果表明,深山含笑叶片总酚超声波辅助提取的最佳提取工艺为：按1：30（质量-体积比）的料液比加入体积分数70％的乙醇,于65℃条件下用超声波辅助提取30min。采用最佳的超声波提取工艺,深山含笑叶片的总酚提取率可达到11．41％。定性分析结果显示,深山含笑叶片的总酚提取物具有典型的酚类化合物特性,并显示出鞣质类成分、黄酮类成分和香豆素类成分的定性反应特征。