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A new fluotescence quenching method has been developed to determine nitrite in water samples. The method is based on the diazo-reaction between nitrite ion and thionine in dilute suifuric acid medium.

文章研究了在稀硫酸介质中亚硝酸根与硫堇发生的重氮化反应,基于硫堇的荧光强度随亚硝酸根的加入量增加而明显降低的现象,建立了荧光猝灭法测定痕量亚硝酸根的新方法。

In this dissertation, firstly, the conditions of isolation and purification of specific toxin fractions produced by Exserohilum turcicum has been studied, and a high-efficient method has been established;secondly, the toxins were analysed by UV-Vis spectrophotometry;then the effect of specific toxin on death rate of corn leaves protoplast was studied by FDA as a tinct reagent;finally, using ANS as a probe and SDS-PAGE to study the membrane protein of the corn leaves with Htl gene. In a word, the aim of the research is to search some existent evidences of the toxin binding site on protoplasmic membrane, and provide proofs for nosogenesis of specific toxin in terms of cytology and the molecular biology.

本研究对玉米大斑病菌1号小种毒素的特异性组分进行分离,建立了高效的特异性毒素分离纯化方法,并对其进行了紫外波谱分析;用荧光素双醋酸酯染色法,测定了毒素特异性组分对原生质体死亡率的影响;进而用1-苯胺基-8-萘磺酸荧光探针标记法和SDS-PAGE电泳法对特异性毒素组分与Htl基因玉米细胞互作过程中的质膜蛋白进行了研究,旨在证明Htl基因玉米细胞质膜上存在特异性毒素组分结合蛋白,为从细胞和分子生物学角度阐明毒素的致病机理提供证据。

METHODS: The BEC were preincubated with crocetin (1, 0.1 and 0.01 μmol/L) for 12 h, then exposed to AGE (100 mg/L). The RAGE mRNA expression was detected by RT-PCR analysis, the intercellular adhesion molecule-1(ICAM-1) was measured by ELISA. The extracellular superoxide ion and thiobarbituric acid reactive substances were assessed with superoxide ion kit and colorimetric assay, respectively The intracellular I2O2 was also detected using the probe 2,7-dichlorofluorescein. The mitochondrial membrane potential and mitochondrial Succinate dehydrogenase were analyzed by the retention of rhodamine 123 (Rh123) and MTT.

不同浓度的西红花酸(1、0.1、0.01μmol/L)预孵BEC细胞12h后,用AGE(100mg/L)刺激细胞12h,RT-PCR法测定RAGEmRNA的表达水平;ELISA法测定细胞间黏附分子-(ICAM-1)的表达;试剂盒分别检测胞外超氧阴离子和硫代巴比妥酸反应产物浓度;同时,还用2,7-二氯荧光素测定了胞内H2O2的浓度,并用罗丹明123(Rh123)荧光法及MTT法分别检测细胞线粒体膜电位水平和其琥珀酸脱氢酶的活性。

The fluorescence images that RhoCr1 coordinated with Cr~(3+)in PC12 cells showed RhoCr1 to be capable of penetrating into living cells and imaging intracellular Cr~(3+)changesFive probes have been designed and synthesized,where different aldehydes were covalently linked to rhodamine B dyes with diethylamine flexible chains.

吸收和荧光光谱的测试结果表明,探针RhoCr1在Tris-HCI中性缓冲溶液中能高选择性的识别Cr~(3+)并产生了荧光增强,通过对探针光谱性能的测试和TEPN滴定实验表明,探针RhoCr1与Cr~(3+)的络合是一个可逆的过程。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank,选择包含人主要T细胞抗原表位序列的人PSCA基因片段,应用异种化抗原设计技术,保留人T细胞抗原表位,设计异种化PSCA融合抗原片段;(2)根据核酸序列按中心模板法设计引物,应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因,以PCR、限制性酶切和DNA序列测定法进行鉴定:(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点,采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4),并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中,转染293T细胞,借助免疫荧光+流式细胞术考察插入片段表达效率,最终选定PSCA_3片段进行下一步研究;(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下,插入pVAX1-neo-IRES—GM/B7载体中,构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB,并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况;(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞,制备小鼠前列腺癌模型,并采用股四头肌肌肉注射+电脉冲法(Electroporation,EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB,同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照,通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率,来评价该DNA疫苗在小鼠体内的抑瘤效果。

MATERIALS AND METHODS: hMSCs labeled with enhanced green fluorescent protein were transplanted into the lateral ventricle of neonatal mouse brain. At 0, 9 and 14 days post-transplantation, MICE were sacrificed and their brains were fixed with 4% paraformaldehyde. The engraftment of transplanted cells in coronal section of the recipient mouse brain was examined wider a fluorescent microscope. Indirected immunofluorescent staining was used to detect the expression of neural proteins of these grafted cells.

材料与方法:将标记绿色荧光蛋白的人骨髓间充质于细胞植入到新生3d的小鼠侧脑室中,分别于移植后0d,9d和14d处死受休鼠,取其脑组织,4%多聚甲醛固定后行冠状面冰冻切片,荧光显微镜下观察hMSCs的植入,激光共聚焦显微镜检测植入细胞神经特异性蛋白的表达,定位双重标记的植入细胞。

These ES cells were introduced into host embryos from outbred KMW though blastocyst injection. Using GFP, the attractive genetic marker, we have been able to follow the fate of ES cells in living embryos and to observe directly the contribution of these cells to mouse embryos. Totally, four GFP-expressing 'green' chimeric mice expressing GFP were obtained.

为研究嵌合体动物中供体胚胎干细胞在宿主胚胎发育中的走向和定位,同时探讨绿色荧光蛋白基因作为报告基因在转基因动物制作中的应用价值,本研究将pEGFP-N1基因导入小鼠ES-D3细胞系,得到稳定表达GFP的胚胎干细胞亚系ES-D3-GFP,通过对昆明小鼠的囊胚腔注射,获得了4只表达绿色荧光蛋白的嵌合体小鼠。

The absorption bands observed for Pb-pheophytin a at 462nm and 673nm , while for Pb-pheophytin b at 480nm and 662nm . Compared with pheophytin a and pheophytin b, all these bands red shifted more or less. Among the reported metallochlorophylls, the degree of red shift of Pb-pheophytin in Soret band was the largest by 54nm and 44nm for Pb-pheophytin a and Pb-pheophytin b, respectively.

实验表明:对铅叶绿素a和b系列来说,它们在Soret带、Q带的特征吸收峰分别在462nm,673nm和480nm,662nm;与脱镁叶绿素a,b相比,铅叶绿素a,b在Soret带和Q带吸收峰均有不同程度的红移;与现已报道的金属叶绿素相比,铅叶绿素a,b在Soret带的红移程度最大,分别为54和44nm;铅叶绿素不具有荧光活性,并能猝灭脱镁叶绿素的荧光。

In order to identify precisely the species of parasites eggs in kimchi and other related products and distinguish from the plant-parasitizing nemotode eggs, we researched the detection technique of Ascaris egg looked as the highest polluting risk of kimchi. The specific fluorescence quantity PCR method was established by using the Ascaris suum genome DNA as positive control,and the optimum way of breaking the eggshells in liquid nitrogen and thawing at 37℃, and the detection method of single egg.

为了精确地对从泡菜及相关产品中发现的寄生虫卵进行种类鉴定,同时与植物本身的线虫卵进行区别,我们在上述研究的基础上,以污染泡菜风险最高的蛔虫卵作为检测目标,进行了模拟阳性泡菜样品中携带蛔虫卵的荧光PCR检测技术研究,即以猪蛔虫基因组DNA为模板,建立了蛔虫特异性荧光PCR检测方法。

Seedlings of Vallisneria natans were planted in waters with different turbidities of 30 NTU, 60 NTU and 90 NTU. Such turbidities were made by mud and sand particles below 100μm in diameter. Photosynthetic fluorescence characteristics were measured by a developed submersible pulse-amplitude modulated fluorometer, Diving-PAM.

用粒径小于100μm的泥沙分别配置浊度为30、60和90NTU的浑浊水体,将苦草幼苗种植于上述水体中,测定幼苗的叶片长和叶片数,并利用水下饱和脉冲荧光仪测定幼苗的光合荧光特性,研究在不同浊度水体中水下光强对苦草幼苗生长的影响。

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But we don't care about Battlegrounds.

但我们并不在乎沙场中的显露。

Ah! don't mention it, the butcher's shop is a horror.

啊!不用提了。提到肉,真是糟透了。

Tristan, I have nowhere to send this letter and no reason to believe you wish to receive it.

Tristan ,我不知道把这信寄到哪里,也不知道你是否想收到它。