染色的

The stem was cut into several segments away from the pine seeding, arranged and collected in the order of chronology time and distance from where they were inoculated. To separate B.xylophilus coarsely out from the stems and made them into paraffin slices. And be observed under the potics microscope by dyed with safranin-fast green contrast-stain method and PI fluorescence stain method. In order to find out how about the nematode rigour distributing , but also ebb and flow of their amounts after invading the host pine seeding. Seek the pathological changing orderliness of host cells in different position along with the remotion of nematodes. By deeply analysing the variety of host cell nucleolus to estimate whether the host cell took place the programmed cell death after inoculated B.xylophilus.

按时间进程和与接种点上下距离的远近，对松苗分茎段进行粗分离和制作成石蜡切片，分别用番红-固绿双重染色法和碘化丙锭荧光染色法在光学显微镜下观察，以了解线虫侵染后在寄主体内的准确分布和数量的消长，寻找随着线虫的移动扩散，寄主不同部位细胞病变的数量、速度发生的变化规律，并进一步通过分析其细胞核的变化来判定松材线虫侵染后是否能诱发寄主细胞发生程序性死亡(programmed cell death，PCD)。

METHODS: RAW264.7 macrophages growth inhibition was measured by MTT assay. The apoptosis effect of RAW264.7 murine macrophages induced by ox-LDL was analyzed by flow cytometric analysis and DNA agarose electrophoresis. After expression of Daxx was silenced by special siRNA, the macrophages apoptosis was observed by AO/EB fluorescence staining. Oil Red O Dyeing experiment was used to show the cellular lipid droplets in lipid-loaded cells. High performance liquid chromatography analysis was performed to determine the content of cellular cholesterol. Real time RT-PCR was used to detect the mRNA expressions of Daxx and SCAP.

用不同浓度的氧化型低密度脂蛋白处理RAW264.7细胞48h，MTT法检测ox-LDL对RAW264.7细胞生长的影响；流式细胞术和DNA断裂片段分析法研究ox-LDL诱导的细胞凋亡；用特异性siRNA沉默Daxx在RAW264.7细胞中的表达，通过AO／EB染色观察基因沉默后的细胞凋亡形态学改变；高效液相色谱检测细胞内胆固醇含量；油红O染色观察细胞内脂滴的形成情况；Real time RT-PCR检测细胞内Daxx mRNA、SCAP mRNA的表达情况；用Western Blot印迹法检测caveolin-1蛋白的表达。

This result suggests that cytomixis occurred at synizesis stage may produce some kind of stress which results in activation of retrotransposon to insert into a new locus, or/and breakage rearrangement at the site of retrotransposon in genomic DNA, that both can be detected by fingerprinning.

分析认为在兰州百合减数分裂前期Ⅰ的凝线期时，由于染色质的胞间运动造成对基因组DNA的胁迫，导致了逆转座子del的激活，所插入的位点序列即为所检测到的特异片段；或者是由于染色质的胞间运动造成染色体的断裂重排。

In this thesis, rat-tails were used as experiment objects, there are 40 slices including transverse and longitudinal, HE-dyed and undyed. Excited by femtosecond laser, the changes of backward SHG and TPEF versus sample preparation, excitation wavelength, excitation power, penetration depth and so on were observed by two-photon laser scanning confocal microscope, effect factors of backward SHG and TPEF and differences of both were discussed and compared, and related comments were also presented.

以鼠尾组织作为实验对象，共40个切片，分为横向和纵向、HE染色和未染色四组，采用飞秒激光器作为激发光，用双光于激光扫描共焦显微镜(two-photon laser scanning confocal microscope， TPLSCM)观察和分析了样品在不同的制备方式、激发波长、激发功率、扫描深度等条件下的背向SHG和TPEF的变化曲线，讨论和比较了生物组织的背向SHG和TPEF的影响因素以及二者之间的异同，并尝试对实验现象做出了一定的解释。

Hyphae, loaded with CTC under pH 6. 8 condition and then grown in pH 8. 0 medium for a little while to destroy the apical acidification, could gave very diffuse fluorescence images of CTC membrame bound Ca〓 and vermiform fluorescence spots due to mitochondrias under pH 6. 8 no longer were clearly distinquished, which implied that besides mitochondrias, other cellular organelles with Ca〓 also were stained with CTC. Thus mitochondrias are not the only one intracellular Ca〓 storage, endoplasmic reticulums and Golgi bodies in the same zone may be also the Ca〓 storages. But the extreme apical zone under 2μm of the growing hyphal tip was still almost devoid of stain under pH 8. 0.The CTC fluorescence was concentrated in the subapic zone beyond about 2μm from the tip and then gradually became lower behind about 40μm from the tip. These results did not support the hypothesis which suggested that the cell wall vesicles in the extreme apical zone were intracellular Ca〓 storages.

将菌丝在pH6.8条件下负载CTC，再置于pH8.0培养介质中短暂生长一会儿以消散菌丝顶端的酸化区域，则CTC膜结合Ca〓呈现弥散的荧光影像，在pH 6.8培养条件下显示的蠕虫状的线粒体荧光斑点不再能够清晰辨认，说明除了线粒体之外，还有其它含Ca〓细胞器，也被CTC染色，线粒体并不是细胞内唯一Ca〓库，还可能包括内质网、高尔基体等细胞器，但菌丝最顶端2μm以前的细胞壁泡囊区域不能被染色，最大荧光强度仍位于菌丝顶端2μm以后区域，约45μm以后荧光变弱，实验结果不支持细胞壁泡囊为菌丝胞内Ca〓库的假设。

The advanced expression of Filaggrin may start the mechanism of apoptosis ahead of tune and the premature expression of TGK may accelerate the formation of cornified cell membrane,which will speed up the terminal differentiation of keratinocytes.Consequently,the hyperproliferative keratinocytes differentiate and exfoliate.

K17在扁平庆皮损中染色为阴性，而在寻常庆、尖锐湿虎皮损中为阳性，这提示：扁平庆的异常增生比寻常庆、尖锐湿庆要弱，这与扁平庆的临床表现和病理改变相一致；K17与炎尖锐湿疣初期表现症的发生有关，在三种庆的病理改变中，扁平庆的炎症现象最微弱，这与K17的染色阴性相吻合。

CML is cytogenetically marked by the philadelphia chromosome, which originates from a reciprocal translocation between chromosome 9 and 22 and is molecularly marked a chimeric bcr-abl gene, resulting from juxta-positive of the abl proto-oncogene on chromosome 9 with the bcr gene, which is normally located on chromosome 22. The chimeric bcr-abl gene expression an 8. 5kb hybrid mRNA transcript giving rise to a 210-KD fusion protein (P210〓) with increased tyrosine kinase activity. P210〓 plays a key role in the pathogenesis of CML. The continuous cell line K562 was established from the pleural effusion of a 53-year-old female with CML in terminal blast crisis, and was a human erythroleukemia line, contained Ph chromosome.

绝大多数慢粒患者白血病细胞中具有Ph染色体，是由9号染色体长臂3区4带和22号染色体长臂1区1带相互易位形成，即t（9；22），使位于9q〓的c-abl原癌基因在第二外显子的5'端断裂并易位到22 q〓的M-bcr基因第2或第3外显子的3'端，形成异常的bcr-abl嵌合基因，该基因转导出异常的mRNA，编码并翻译出P210蛋白，该蛋白具有很强的酪氨酸激酶活性，使粒细胞发生恶性增殖。K562细胞属于慢粒急变、红白血病细胞株，具有Ph染色体。

Methods: A model to evaluate lymphocyte proliferation stimulated with a polyclonal activator, concanavalin A, was established by vital dye carboxyl fluorescin diacetate succinmidyl ester labeling technique. Effects of the different doses of anisomycin on the lymphocyte proliferation were estimated by flow cytometry and MTT methods. The propidium iodide labeling technique was applied to assay the effect of the different doses of anisomycin on changes of the lymphocyte cell-cycle stimulated by ConA or by phorbol ester plus Ionomycin. The percentage of the expression level of CD69 and CD25 on the activated lymphocytes was evaluated by fluorescin-conjugated monoclonal antibody double labeling technique.

以活体染料羧基荧光素乙酰乙酸琥珀酰亚胺酯染色，建立在多克隆刺激剂刀豆蛋白A刺激下评价小鼠淋巴细胞增殖的模型，通过流式细胞术和MTT法分析茴香霉素在不同剂量下对淋巴细胞增殖的作用；采用碘化丙锭染色分析茴香霉素对ConA或佛波醇酯加离子霉素刺激的小鼠淋巴细胞周期变化的作用；利用荧光标记的单克隆抗体双染技术和流式细胞术观察茴香霉素对小鼠CD3~+T细胞早期及中期活化标志分子CD69和CD25表达的影响。

The biomechanical tests showed that two kinds of artificial bones had not significant difference on compressive strength and Young\'s modulus(P＞0.05),while the flexural strength of nano-nacre artificial bone was less than the control group(P＜0.05).3.The results of CCK-8 showed that the difference were not significant in each group,the proliferation of osteoblast reached the peak at the 5th day;7 days after being co-cultured,the total protein content of study group was higher than control group and blank group(P＜0.05),while the difference between control group and blank group was not significantP＞0.05The difference of alkaline phosphatase activities among three groups was not significant(P＞0.05The SEM view showed that osteoblast attached and grew well in two kinds of artificial bone.4.X-ray photography showed that two kinds of powder started to degrade in 2 weeks;this phenomenon became more appear in 4 weeks,nano-nacre powder degraded faster than micron-nacre powder,while the hole shadow was easy to be found;in 8 weeks,all the femoral holes recovered and returned to normal bone mineral density in all groups.Analysis of tetracycline fluorescent double marks in the hard tissue grinding slices indicated that new bone grew fastest around the bone defect area in study group,while most slowly in blank groupP＜0.05 SEM(scanning electron microscope observation showed that nano-nacre powder degraded more quickly.The same result can be found through the demineralized sections morphometric analysis,and both of the composite artificial bones made from those two kinds of nacre powder had the good connection with the adjacent tissue in rats body without apparent inflammatory response.5.X-ray photography showed that rabbit\'s bone defects healed faster in study group since NNAB implanted than in control group since MNAB implanted.At 24 weeks after operation,bone density in radial defects had nearly accessed to the normal area,while lower in control group,and turned up nonunion in blank group;The checking of BMD showed that results in study group were higher than those in control group at 8,16 and 24 week(P＜0.05), and the difference between the BMD values in study group at 24 week and those in blank group was not significant(P＞0.05).The gross specimens showed satisfactory histocompatibility both in study group and in control group,with bone tissue growing from two sides into the center of implanted materials; Normal slices in HE stain and hard tissue grinding slices in Stevenel\'s blue/Van Geison\'s picro-fuchsin stain showed that the bone growth tendency was better in study group than that in control group,and the medullary cavity had been penetrated to the implanted materials in study group at 24 week;Analysis of tetracycline fluorescent double marks in the hard tissue grinding slices indicated that new bone in both groups grew fastest 8 weeks after surgery,while slow down at 16 week.

纳米珍珠层/消旋聚乳酸复合人工骨与微米珍珠层/消旋聚乳酸复合人工骨分别与成骨细胞共培养后，其各时间点CCK-8法检测值与空白对照无显著差异(P＞0.05)，成骨细胞均在第5天达到增殖高峰期；培养7天后，实验组细胞蛋白含量高于对照组及空白组(P＜0.05)，后两者之间则无显著差异P＞0.05碱性磷酸酶活性在三组间均无显著差异(P＞0.05电镜下可见成骨细胞在两种人工骨上都有良好生长贴附能力。4.X-ray显示两种粉体在大鼠股骨骨洞植入第2周时都开始出现了降解，第4周时更为明显，纳米珍珠层粉较之微米珍珠层粉降解更快，而空白对照组骨洞阴影仍可见，至8周时，则所有组骨洞均己闭合修复，X-ray下已不可见原钻孔痕迹，恢复正常骨质密度；硬组织磨片四环素荧光双标记结果显示纳米珍珠层粉植入组较其余两组在骨缺损区周围新骨生长速度更快，空白组速度最慢P＜0.05电镜观察及常规脱钙切片亦可见到纳米粉体降解较快；由以上两种原材料制得的纳米珍珠层/消旋聚乳酸复合人工骨与微米珍珠层/消旋聚乳酸复合人工骨在大鼠体内均与周围组织结合良好，无明显炎症反应。5.X-ray显示纳米珍珠层/消旋聚乳酸复合人工骨植入兔桡骨缺损区后其骨愈合速度较对照组微米珍珠层/消旋聚乳酸复合人工骨植入的快，至植入术后24周，实验组骨缺损区接近正常骨密度，对照组骨缺损区密度较低，空白组则呈现骨不连状态；骨密度测量结果显示术后8周、16周、24周实验组的骨密度值高于对照组(P＜0.05，24周实验组的骨密度值与术前所测得的正常值无显著性差异P＞0.05动物取材大体所见均显示组织相容性良好，骨组织逐渐由植入材料两端向中央生长；常规切片HE染色及硬组织磨片Stevenel\'s blue/Van Geison\'s picro-fuchsin联合染色均可见实验组骨缺损区长势优于对照组，至术后24周，实验组骨髓腔与材料已呈相交通状；硬组织磨片荧光显微镜下观察，两组材料在术后8周处于骨生长最快速时期，16周时速度开始减慢，术后4、8、16周时实验组的新骨生长速度均较对照组的快

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 labia have now been sutured together almost completely.The drains and the Foley catheter come out at the top.

此刻阴唇已经几乎完全的缝在一起了，排除多余淤血体液的管子和Foley导管从顶端冒出来。

To get the business done, I suggest we split the difference in price.

为了做成这笔生意，我建议我们在价格上大家各让一半。