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Methods: A 5ml bone marrow was extracted from the lilac of human volunteers. By Percoll fluid and density gradient centrifugation, the MSC was obtained; after the cells filled the bottom of vessel, subcultured them, when they subculture in third generation, redigested them, 500 R/min centrifugate, alter the completed medium to chemical definition medium, examined the form change and prolifration of cells by invert microscope, toluidine blue stain、immunocytochemical stain and RT-PCR to test the type Ⅱ collagen mRNA and proteoglycan.

取健康成人髂后上棘处骨髓5ml,经percoll液分离后密度梯度离心,〓/ml密度接种培养,观察原代细胞的贴壁、增殖状况,细胞长满瓶底后进行传代培养;传至第三代细胞,重新消化后以500转/分钟轻度离心5分钟,〓/ml接种,改用化学限定培养基代替完全培养基培养,倒置显微镜观察细胞生长情况及形态变化,甲苯胺蓝染色观察诱导细胞合成细胞外基质中的蛋白多糖,免疫细胞化学染色检测ECM中Ⅱ胶原的蛋白合成,RT-PCR鉴定诱导细胞Ⅱ胶原mRNA的表达。

RESULTS: At 16 weeks after the compound of after-induced chondrogenetic BMSCs and SIS were transplanted, the materials had been completely absorbed and there were new cambium in the experimental group, which proved to be chondrocytes after hematoxylin-eosin stain, Massan stain and Toluidine blue dyeing.

结果:诱导软骨细胞-小肠黏膜下层复合物在植入体内16周时材料明显被吸收完全,且有软骨生成,与正常组织未见明显区别,但修复层软骨较薄。经苏木精-伊红染色,Massan染色,甲苯胺蓝染色检查证实为软骨组织。

The color of elytral stain is an important morphological character traditionally distinguishing Anoplophora glabripennis from A. nobilis. The typical stain color of A. glabripennis is white, but that of A. nobilis is yellow.

鞘翅上斑纹的颜色是传统形态分类区分光肩星天牛和黄斑星天牛的重要外部形态特征,光肩星天牛的鞘翅斑纹颜色多为乳白色,而黄斑星天牛则为黄色。

PartⅡ: Analysis of the paraffin embeded tissue slides [obtained from trail and control group at pre-treatment (d0) and post-operation (d15±1 days) ] and frozen section slides (from 4 typicalcases: 1 of IAC group, the other sBRM group at d5, d7 and d10), by H-E stain, monoclonal antibodies (CD3/CD4/CD8/CD68/CD57) marking, SP-DAP or APAAP-AEC stain, and TUNEL in situ apoptotic cell DNA fragments fluorescin labeling.

二。组织病理学研究:对上述两组病人治疗前后原发灶肿瘤组织蜡块分别作H-E和CD3、CD68表型免疫组化染色;并对其中四例典型病人的疗中间断连续活检新鲜组织作H-E染色,CD3、CD4、CD8和CD57表型免疫组化染色和凋亡细胞DNA断片TUNEL法荧光标记检测。

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 results showed that at the beginning stage of being attacked by Chinesis white pine beetle and their symbiotic blue-stain fungus, PAL, PPO, SOD activities and MDA content significantly increased in the phloem of P. armandi, and decreased with the increase of the number of D. armandi and the attacking on the xylem and phloem of P. armandi from blue-stain fungus, then reached the minimum level in withering stage of P. armandi. But the contents of nutrition materials in the phloem, such as water, protein, glucide, fat, amylum and so on, declined with the infestation of D. armandi and symbiotic fungus. In summary, the resistance of physiology and biochemistry in the phloem of P.

结果表明,在华山松大小蠹和共生蓝变真菌危害初期,华山松韧皮部内的苯丙氨酸解氨酶、多酚氧化酶、超氧化物歧化酶活性均显著提高,丙二醛含量明显增加;但随着华山松大小蠹种群数量的增加和蓝变真菌对华山松韧皮部和木质部组织危害的加剧,韧皮部组织内的PAL、PPO和SOD活性逐渐降低,在枯立木阶段降至最低;与此同时,华山松韧皮部内的水分及蛋白质、糖类、粗脂肪、淀粉等营养物质含量,则随华山松大小蠹和共生蓝变真菌危害的加剧而不断减少。

Results Among the three groups,the children's rib cartilage had the most blood vessels,the most chondrocytes,well-distributed stain of matrixes,and the type Ⅱ collagen was expressed actively and highest in photedensity.The rib cartilage of teenager group had less blood vessels,unhomogeny distributed stain of matrixes,the enlarged and separated cartilage lacunas.The rib cartilage in adult group showed the least blood vessels,the least chondrocytes.the hyalinization of perichondium,the most deposition of calcium salt,and the type II collagen was expressed at the lowest level in photodensity.

结果 儿童组肋软骨膜血管最丰富,软骨基质染色均匀,软骨细胞数目最多,Ⅱ型胶原蛋白表达最活跃,平均积分光密度值最高;青少年组软骨膜内血管减少,软骨基质染色出现明显的不均质状,软骨陷窝体积变大,并呈分隔状,陷窝内软骨细胞数目减少,II型胶原蛋白表达较儿童组减弱;成人组软骨膜血管、细胞成分明显减少,软骨膜内的纤维成分明显玻璃样变,钙盐沉积较青少年组时明显增多,Ⅱ型胶原蛋白表达较青少年组减弱。

At the end of 12 and 18 weeks, rats were killed, taken the muscle of posterior limb, after separating the gastroenemius、soleus、plantaris, to prepare the frozen section of the each kind of muscle respectively to be confirmed to form the myopathy model by the histochemical stain.(3) RT-PCR、Western blot and immunohisto-chemistry stain were respectively used to detect the expression of NF-κB、MMP-2 and MMP-9 during different period.

2实验组用酒精给大鼠灌胃,对照组用含与酒精体积相等的生理盐水灌胃,两组均喂饲改良的全营养高脂饲料,于12周及18周后,取后肢肌肉,在分离出腓肠肌、比目鱼肌、跖肌后,将各类肌肉均分别地制备成冰冻切片和石蜡切片,经组织化学染色确认成模。

Methods KC and FE of tympanic membrane were cultured on the surface of CCM prepared by torrefaction, then the cells biological characteristics were detected by immunofluorescence stain of keratin and vimentin, and the cells proliferative activity were identified by immunofluorescence stain of PCNA.

方法热干法制备CCM,传代的大鼠鼓膜上皮KC和FB分别接种于培养板及CCM表面,通过角蛋白、细胞膜波形蛋白免疫荧光染色鉴定细胞特异性,通过细胞核增殖抗原(proliferating Cell nuclear antigen, PCNA)免疫荧光染色测定膜上细胞的增殖能力,通过细胞培养液上清的经脯氨酸测定评价对FB胶原分泌功能的影响。

Five rats from both groups were killed at days 7,14 and 28,respectively.And we used deproteinization analysis to detect the plasma H2S concentration and hematoxylin and eosin stain and Mallory trichrome stain for the lung tissue.

观察指标:于试验的第7、14、28天时每组分别处死5只大鼠,采用去蛋白分析方法测量血浆中H2S含量,取肺组织行HE染色和Mallory三色染色。

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