染色法

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

HepG2 cells were exposured to oxymatrine, final concentrations of which were 0.1, 0.2, 0.3, 0.4, 0.5, 0.8, 1, 2, 5, 8, 10, 15 g.L-1 respectively, MTT method was used to determine the inhibiting effects of oxymatrine on HepG2 cells proliferation, followed by Haematoxylin and Eosin staining to observe the cell morphology.

以0.1，0.2，0.3，0.4，0.5，0.8，1，2，5，8，10，15 g.L-1的氧化苦参碱处理HepG2细胞，四噻唑蓝法检测其对细胞增殖的影响；苏木精-伊红染色比较细胞形态差异；免疫组化染色进一步检测HepG2细胞肿瘤相关蛋白表达量的变化。

Ten adult mixed breed dogs without significant individual difference were made transverse fracture at bilateral iliopectineal crest 1.5cm above the dome of acetabulum, which were fixed with ATMF'S or steel plate respectively. The animals were sacrificed and specimens were procured at 2, 4, 6, 8 and 12 weeks after operation. Samples from the fracture gaps were investigated by histology of HE and Masson staining, image pattern analysis of new bone formation.

方法］ 选用10只成年杂种家犬，双侧髋臼臼顶上方1.5cm处横形截骨，分别采用ATMFS（骨盆髋臼三维记忆内固定系统）前柱固定器和5孔重建钢板内固定，于术后2、4、6、8、12周处死动物取材，行HE染色、Masson三色法染色组织学检查，图像分析新骨形成积分光密度值，研究不同应力作用模式下的骨盆骨折愈合情况。

Method］Ten adult mixed breed dogs without significant individual difference were made transverse fracture at bilateral iliopectineal crest 1.5 cm above the dome of acetabulum, which were fixed with ATMFS or steel plate respectively. The animals were sacrificed and specimens were procured at 2,4,6,8 and 12 weeks after operation.Samples from the fracture gaps were investigated by histology of HE and Masson staining,image pattern analysis of new bone formation.

方法］选用10只成年杂种家犬，双侧髋臼臼顶上方1.5 cm处横形截骨，分别采用ATMFS(骨盆髋臼三维记忆内固定系统)前柱固定器和5孔重建钢板内固定，于术后2、4、6、8、12周处死动物取材，行HE染色、Masson三色法染色组织学检查，图像分析新骨形成积分光密度值，研究不同应力作用模式下的骨盆骨折愈合情况。

Method］Ten adult mixed breed dogs without significant individual difference were made transverse fracture at bilateral iliopectineal crest 1.5 cm above the dome of acetabulum, which were fixed with ATMFS or steel plate respectively. The animals were sacrificed and specimens were procured at 2,4,6,8 and 12 weeks after operation.

］选用10只成年杂种家犬，双侧髋臼臼顶上方1.5 cm处横形截骨，分别采用ATMFS(骨盆髋臼三维记忆内固定系统)前柱固定器和5孔重建钢板内固定，于术后2、4、6、8、12周处死动物取材，行HE染色、Masson三色法染色组织学检查，图像分析新骨形成积分光密度值，研究不同应力作用模式下的骨盆骨折愈合情况。

［Objective］To explore the effect of continuous physiological osteogenic stress on the bone healing of canine iliopectineal crest.［Method］Ten adult mixed breed dogs without significant individual difference were made transverse fracture at bilateral iliopectineal crest 1.5 cm above the dome of acetabulum,which were fixed with ATMFS or steel plate respectively. The animals were sacrificed and specimens were procured at 2,4,6,8 and 12 weeks after operation.Samples from the fracture gaps were investigated by histology of HE and Masson staining,image pattern analysis of new bone formation.

探讨持续顺应生理力线的压应力对犬骨盆弓状线部骨折愈合的影响［方法］选用10只成年杂家犬，双侧髋臼臼顶上方1.5 cm处横形截骨，分别采用ATMFS(骨盆髋臼三维记忆内固定系统)柱固定器和5孔重建钢板内固定，于术后2、4、6、8、12周处死动物取材，行HE染色、Masson三色法染色组织学检查，图像分析新骨形成积分光密度值，研究不同应力作用模式下的骨盆骨折愈合情况。

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 cartilage histological characteristics were observed according to the method of Mankin. Immunohistochemical staining was performed to investigate the expression of VEGF.

按照Hulth法建立膝骨关节炎模型，术后第5周起至第7周末，阳和汤组予混有阳和汤的颗粒饲料，正常组及模型组均给予普通饲料；术后第8周分别取各组胫骨平台关节软骨，采用番红O染色，进行Mankin评分；免疫组织化学染色测定关节软骨中VEGF表达的阳性指数。

Light microscope and transmission electron microscopy showed that SMMC-7721 cells induced by SAHA had undergone the restorational alteration in morphology and ultrastructure, which were different from those of nontreated cells but were similar to those of normal cells, and the changes were as follows: the cells turned to be flat and spread; the nucleo-cytoplasmic ratio lessened and nuclear shape became rather regular; the number of nucleolus reduced and its volume lessened; euchromatin increased while heterochromatin decreased in nucleus; in the cytoplasm, mitochondria grew in number with relatively consistent structure and well-developed mitochondria cristae; Golgi complex turned to be well-developed and typical; rough endoplasmic reticulum increased. Immunocytochemistry assay showed that the expression of AFP and PCNA were declined significantly. FCM analysis showed SAHA could arrest SMMC-7721 cells in G0/G1 phase, with an accumulation of the cells in G0/G1 phase while a decrease of cells in S phase. Semi-quantitative RT-PCR detection revealed that the expression of p21WAFl mRNA was upregulated remarkably in the cells treated with SAHA.

结果：倒置显微镜和透射电镜观察显示，经SAHA处理的细胞增殖速度显著减慢，细胞体积增大，细胞核较小，形状较为规则，核仁数量减少、体积变小，核内常染色质增多而异染色质减少，核质比例减小，细胞质内线粒体数量增多、线粒体嵴发达，高尔基体较为典型，粗糙型内质网增多，呈现出与正常上皮细胞相似的形态变化；MTT比色法测定结果显示不同浓度(2.5、5.0、7.5、10.0uM)SAHA对SMMC-7721细胞的增殖均有抑制作用，并有明显的剂量依赖和时间依赖关系；免疫细胞化学检测显示SAHA能显著降低PCNA和AFP在SMMC-7721细胞中的表达；流式细胞仪检测结果显示，SMMC-7721细胞经SAHA处理后，G0/G1期细胞明显增加，S期细胞则明显减少，细胞被阻滞于G0/G1期；RT-PCR检测结果表明，SAHA作用12h后SMMC-7721细胞中p21WAF1 mRNA的表达即有增加，24h后更为明显。

AMs that collected, pured and cultured with contine method were stimulated by LPS of different concentration(0μg/ml,0.01μg/ml,0.1μg/ml,1μg/ml,10μg/ml) for 60min or by 1μg/ml LPS for different time stage (0min,5min,15min,30min,60min,120min) to observe the dynamic change of NF-кB intranuclear level and NO production, from which the best concentration and time point of LPS stimulation were selected. In the study, all AMs were divided into 4 groups: control group, group stimulated with LPS, group interrupted by Cal C and group inhibited by PDTC. The following parameters were measured: NF-кB level in nuclear protein extraction of AMs detected with sandwich ELISA, Inter-nuclear transposition of NF-кB observed with immunocytochemistry staining, NO content in cell culture medium quantitied with nitric acid reductase assay, Morphologic change of AMs in apoptosis observed with acridine orange staining and fragmentation at genome DNA of AMs detected with apoptotic electrophoresis assay.

分离、纯化及培养大鼠肺巨噬细胞；以不同浓度的LPS(0μg/ml，0.01μg/ml，0.1μg/ml，1μg/ml，10μg/ml)和不同作用时间(0min，5min，15min，30min，60min，120min)分别刺激小室培养的细胞单层，观察NF-κB的核内浓度及NO合成量的动态变化，选择LPS的最佳用量和作用时间；然后分成四组实验，设正常对照组，LPS处理组，特异性PKC抑制剂阻断组，NF-κB抑制剂阻断组；收集培养的单层细胞及培养液；采用夹心ELISA法定量测定细胞核提取物中的NF-κB水平；免疫组化法检测NF-κB的核内移位变化；硝酸还原酶法测定细胞培养液中NO含量；吖啶橙染色观察凋亡细胞的形态学变化，凋亡电泳实验检测细胞凋亡后基因组DNA的断裂情况。

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。