结晶紫

Methods Anhydrous acetic acid as solvent, 0.1 mol/L perchloric acid as titrant, crystal violet solution as indicator to establish non-aqueous titrations and ODS column as stationary phase, methanol and a mixture of 0.025 mol/L sodium phosphate monobasic and 0.02 mol/L sodium dedecyl sulfate (80:20) adjusted with phosphoric acid to a pH (4.8±0.1J as mobile phase, 220 nm as detective wavelength to establish HPLC-UV method for determination of piperazinylethylestrone drug substance; FMOC-CL as a derivatization reagent, FD as a detector to establish an HPLC method with derivatization and column switching for determination of piperazinylethylestrone in dog plasma.

分别采用无水冰醋酸为溶剂，0.1mol/L高氯酸为滴定液，结晶紫指示液指示终点的非水滴定法和HPLC-UV法测定哌嗪雌酚酮原料药含量；以FMOC-CL为衍生化试剂，采用柱切换柱前衍生化萤光检测的HPLC法测定狗血浆中哌嗪雌酚酮含量。

Adsorpytion of congo red and crystal violet can be described using the Langmuir isotherm, on the other hand, adsorption of methylene blue and aniline blue can be described using a Freundlich isotherm.

对刚果红和结晶紫的吸附服从Langumir吸附等温线，而对于亚甲基蓝和苯胺蓝的吸附则服从Freundlich吸附等温线，图4，表2，参9。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法，从人组织细胞总RNA中扩增可溶性TWEAK胞外区（sTWEAK1和sTWEAK2）的cDNA序列及全长编码序列，用琼脂糖凝胶电泳分析PCR产物，胶回收目的基因片段，连接到pMD18-T克隆载体中，转化大肠杆菌K802，PCR和酶切筛选阳性克隆，全自动DNA测序验证序列；(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中，分别转化大肠杆菌BL21和TB1，PCR筛选和酶切鉴定，阳性克隆用IPTG诱导表达，表达产物用SDS-PAGE分析和Western blot验证融合蛋白；(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白；(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测，贴壁细胞用结晶紫染色法，悬浮细胞用磺酰罗丹明B染色法，酶标仪检测OD值；(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量；(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况；(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响；(8)用双荧光素酶报告基因检测法，测定表达产物对敏感细胞NF-κB的影响；(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上，用脂质体转染法转染HEK293细胞，PCR鉴定重组质粒。

METHODS: Using tissue culture technique, the maximal non-toxic concentration of luteolin in swine testicle cell culture was measured, and the inhibitory effect of luteolin on TGEV was measured by cytophathogenic effect observation and crystal violet measurement. In addition, the degrees of inhibition of the luteolin on CPE of virus in cell culture were observed by changing sequence of adding the vires and the drag. RESULTS : The maximal non-toxic concentration of luteolin was 873.39 μmol/L.

采用细胞培养法，通过结晶紫摄入量测定结合细胞病变作用，观察木犀草素在不同的加药方式下，对感染TGEV的猪睾丸细胞的保护作用、结果：TGEV病毒先吸附ST细胞再加入药液，先与TGEV病毒孵育再接种于卯细胞，或药液先与细胞孵育再用病毒感染等不同方式下，木犀草素在0.84-54.50μmaol／L浓度范围内，均能明显地提高细胞的存活数。

The characteristic property applying the matter absorption spectrum , thing optics characteristic property having studied crystal is purple under different condition and the purple ion that anionic surfactants twelve alkyls mahogany acid sodium forms association.

应用物质的吸收光谱特性，研究了不同的条件下结晶紫与阴离子表面活性剂十二烷基磺酸钠形成的紫色离子缔合物的光学特性。

Part III The Effect of Indomethacin on Hyphal Formation of Candida Albicans and Its Mechanism Research Experiment one The Effect of Indomethacin on the Hypha Formation in the Liquid Medium In this experiment, to observe the hypha formation of three strain candida albicans after different concentration Indomethacin was added in RPMI1640 contained 10% calf serum in microscope (x400), and to analysis the percent rate of hypha formation with crystal-violet germ tube assay.

第三部分消炎痛对白色念珠菌菌丝形成的影响及其机制探讨实验一在液体培养基中消炎痛对菌丝形成的影响本实验主要研究在含10%小牛血清的RPMI1640液体培养基中同时加入不同浓度消炎痛后，分别在2.5h后和4h后在显微镜下(x400)观察三株白色念珠菌芽管、菌丝形成情况，并用结晶紫芽管检测法定量分析菌丝形成的百分率。

Experiment three The Effect of Indomethacin on Hypha Formation when Exogenous PGE_2 Is Present In this experiment, to study the effect of Indomethacin on hypha formation after Indomethacin (2.0mM) and exogenous PGE_2 (300pg/ml) were added in RPMI1640 with crystal violet-based germ tube assay and to further interpret the mechanism of Indomethacin inhibiting hypha formation.

实验三在外源性PGE_2存在时消炎痛对菌丝形成的影响本实验主要研究在RPMI1640培养基中同时加入外源性PGE_2(300pg/ml)和消炎痛(2.0mM)，培养4h后用结晶紫芽管检测法定量分析消炎痛对菌丝形成的影响，以期能进一步说明消炎痛抑制菌丝形成的作用机制。

With hydroxylamine hydrochloride as reducing agent mixed gold chloride acid on the surface of the second deposit silicon film growth, measured by scanning electron microscopy of the surface morphology of silicon, this new system of surface-enhanced Raman scattering activity the study of crystal violet probe molecules to study the SERS activity revealed that the system is a very efficient SERS substrate activity, and the SERS activity is better than chemically prepared gold plastic.

用盐酸羟胺作为还原剂混合氯金酸对硅片表面进行二次金膜的生长，用扫描电子显微镜测定硅片的表面形貌，对这种新体系的表面增强拉曼散射活性进行了研究，以结晶紫为探针分子，研究其SERS活性发现该体系是一种非常高效的SERS活性增强基底，并且其SERS活性优于用化学方法制备的金胶。

With hydroxylamine hydrochloride as reducing agent mixed gold chloride acid on the surface of the second deposit silicon film growth, measured by scanning electron microscopy surface morphology of gold films, this new system of surface-enhanced Raman scattering activity the study of crystal violet probe molecules to study the SERS activity revealed that the system is a very efficient SERS substrate activity, and the SERS activity is better than chemically prepared gold plastic.

用盐酸羟胺作为还原剂混合氯金酸对硅片表面进行二次金膜的生长，用扫描电子显微镜测定金膜的表面形貌，对这种新体系的表面增强拉曼散射活性进行了研究，以结晶紫为探针分子，研究其SERS活性发现该体系是一种非常高效的SERS活性增强基底，并且其SERS活性优于用化学方法制备的金胶。

Treatment can be 1%-2% sodium bicarbonate solution, pre-cleaner mouth in the breast-feeding or nystatin (10 million units / ml) coated surface of the skin 3-4 times a day; or coated with 1% crystal violet solution surface of the skin 2-3 times a day.

治疗可以是1％-2％碳酸氢钠溶液，预清洗口哺乳或制霉菌素（1000万单位/毫升）涂在皮肤表面，每天3-4次，或用1％结晶紫溶液涂在皮肤表面，每日2-3次。

This one mode pays close attention to network credence foundation of the businessman very much.

这一模式非常关注商人的网络信用基础。

Cell morphology of bacterial ghost of Pasteurella multocida was observed by scanning electron microscopy and inactivation ratio was estimated by CFU analysi.

扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。