荧光镜的

Electron donating substituent CH〓, OCH〓 and 4, 5-benzo make λ〓 red shift, electron with -drawing substituent F, Cl and CF〓 make λ〓 hypsochromic shift; b The W〓 is bigger than relative symmetrical carbocyanines, and the ratio of 0-1＇ and 0-0＇ transition intensity increase with the increase of σ〓; c The absorption and the fluorescence spectra are mirror image, and the λ〓 have a good linear relationship; d The Stokes shift △λ〓 and the Hammett σ〓 constant have a good linear relationship.

研究了染料的光物理性质，①苯环上取代基影响染料的λ〓和摩尔消光系数，相对于H，给电子取代基CH〓、OCH〓、4，5-benzo使λ〓红移，摩尔消光系数减小，吸电子取代基F、Cl、CF〓使λ〓蓝移，摩尔消光系数减小；②化合物吸收峰的半峰宽较相应的对称染料的大，且振动肩部与最大吸收峰的强度比随σ〓增大而增大；③吸收光谱与荧光光谱呈镜像关系，且λ〓与λ〓呈良好的线性关系；④化合物的Stokes位移△λ〓与取代基的Hammett σ〓常数呈线性关系。

The expression changes of relating-apoptosis gene proteins (bcl-2, bax) were detected by immunohistochemistry before and after treating with the Agaricus Blazei Murill extract to explore the possible mechanisms of inducing apoptosis for MGC-803 cells in vitro. Results: The Agaricus Blazei Murill extract significantly inhibited the proliferation of gastric cancer cells in vitro and the inhibitive effects were depended on the medicine concentration and treating times. After treating 24 hours on the gastric cancer cells with the morphologic changes of apoptosis with chromatin margination, karyopyknosis, karyorrhexis were found by the light.

结果：(1)通过细胞培养和MTT法，表明长白山姬松茸在体外对MGC-803细胞株有明显的抑制作用，加药组与对照组相比其生长抑制率有显著性差异(P＜0.01)，而且这种抑制作用呈现浓度和时间的依赖性；(2)通过集落形成率的测定结果表明，加药组与对照组相比其集落形成率和集落形成抑制率有明显差异(P＜0.01)，说明长白山姬松茸对MGC-803胃癌细胞株有明显抗增殖作用；(3)光镜观察结果表明，加药组可见细胞脱水浓缩伊红染色增强，胞体缩小，内含高度浓缩的胞核呈深蓝色等典型细胞凋亡形态；经AO／EB荧光染色观察结果表明，当终浓度为1.0mg／ml的姬松茸提取物作用于MGC-803胃癌细胞24h后，其凋亡率和破膜率与自然凋亡率与破膜率相比，均有显著性差异，其凋亡率86.3％(P＜0.001)，破膜率为41.6％(P＜0.01)，说明姬松茸确实有诱导MGC-803胃癌细胞凋亡的作用，同时也有细胞毒作用，但以诱导细胞凋亡为主；(4)免疫组化结果表明，用药前后凋亡相关基因的BCl-2、Bax蛋白均有显著性的改变(P＜0.001)。

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。

Second is studying the traumatic condition changes in a choosey pressure at different post-PT phases. Moreover, we detected model PLA_2 activity, express of PLA_2 subtype mRNA by fluorescent quantitative RT-PCR technique, protein express and site by Western blot and IHC at different post-PT phases. At last, we studied the PLA_2 inhibited effects and model protection of Variabilin in vivo.

研究在不同压强的外力作用下，模型指定时相点的一般情况和存活率、大体、光镜及电镜下胰腺组织的病理变化和常规酶学变化，得到稳定建模致伤压强；然后，固定致伤压强，研究胰腺创伤后不同时相点的伤情变化(包括一般情况、胰腺组织的病理变化、AMS和LPS活性变化、血常规、血生化、胰腺细胞凋亡以及胰腺组织细胞生长周期变化情况)；再次，采用3H-花生四烯酸标记大肠杆菌膜作为底物的方法检测大鼠胰腺创伤后不同时相点的PLA_2活性，利用荧光定量RT-PCR技术检测其亚型mRNA的表达，利用Western blot和IHC技术检测其蛋白表达量和蛋白组织定位；最后，将Variabilin体内应用于急性胰腺创伤大鼠模型，研究其对模型的保护作用和对PLA_2的抑制效应。

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。