染色法

Methods: A sepsis model of mice made with cecum deligation and perforation was established,and a radioimmunoassay for FABP and 96-well spectrophotometry assays for myeloperoxidase and superoxide dismutasewhich were related with clearance of free radicals,were used to detect their levels in lung and intestine ho...

建立小鼠盲肠结扎致脓毒症模型，采用放射免疫分析法和96孔分光光度分析法分别检测肺肠组织匀浆液中FABP及髓过氧化物酶、超氧化物歧化酶等与自由基清除相关的酶的水平、同时以HE染色方法观察肺肠组织病理组织学改变。

Real-time quantitative pcr, immunohistochemistry, dry-wet weight method, histological techniques and haematoxylin and eosin stain were used to detect expression of icam-1, the change of mrna at different time phases, water containing in the brain tissue and the inflammatory infiltration respectively after diffuse brain injury.

采用marmarou方法获得大鼠弥漫性脑损伤模型，实时定量rt-pcr、s-p免疫组化法分别测定icam-1蛋白和mrna在外伤后不同时间点的表达变化，干湿重法测脑组织含水量，组织切片苏木精-伊红染色观测炎性细胞浸润情况。

Methods The rat models of diffuse brain injury were established by Marmarou's method. Real-time quantitative PCR, immunohistochemistry, dry-wet weight method, histological techniques and haematoxylin and eosin stain were used to detect expression of ICAM-1, the change of mRNA at different time phases, water containing in the brain tissue and the inflammatory infiltration respectively after diffuse brain injury.

采用Marmarou方法获得大鼠弥漫性脑损伤模型，实时定量RT-PCR、S-P免疫组化法分别测定ICAM-1蛋白和mRNA在外伤后不同时间点的表达变化，干湿重法测脑组织含水量，组织切片苏木精-伊红染色观测炎性细胞浸润情况。

Image analysis system was used for semi-quantitive assay of TGF-βRⅠ and R Ⅱexpression, and microphotography was used to record the morphologic changes of culturedchondrocytes. The results are:1. TGF-〓 showed sequentially inhibitory and promotive effects on the proliferation of human articular chondrocytes of two week monolayer culture from one passage to eight passage.

本研究采用CellTiter 96〓单溶液细胞增殖分析试剂盒测定体外培养中的软骨细胞的增殖活细胞数量；免疫组化法测定软骨细胞TGF-βR Ⅰ、R Ⅱ，Ⅱ型胶原和S-100蛋白的定性表达；酶免分析法测定TGF-〓作用后的Ⅱ型胶原定量表达；通过图象分析仪，对TGF-βRⅠ、R Ⅱ的免疫组化染色结果进行半定量分析；本研究的实验结果如下：1。

METHODS: NSCs were isolated from neonatal rats by enzyme digestion and mechanical separation. At the fourth passage, cell clone masses received nestin immunocytochemistry. Remaining cells were dispersed by mechanical separation. Monoclone NSCs were incubated by limiting dilution assay, and made into 108 L-1 monoplast suspension in complete medium. NSCs were assigned into 2 groups. NSCs in the control group were incubated in 10% fetal bovine serum.

酶消化和机械分离法相结合体外分离培养新生鼠神经干细胞，传至第4代的细胞克隆团行巢蛋白免疫细胞化学染色观察，剩余细胞团用机械法分散，采用有限稀释法进行单克隆神经干细胞培养，加入完全培养基制成108 L-1的单细胞悬液，分为2组滴入培养板，对照组加入10%FBS，诱导组加入10%FBS+50 μg/L神经生长因子，培养5~7 d。

The immunocytochemistry was used to identify the RASMC.RESULTS: The RASMC were successfully cultured by the adherent method of tissue explants and grown in the "peak- valley" mode. The cultured RASMC were fusiform shape with a big oviform or round nuclei rich in cytoplasm. Immunohistochemical method ( S-P method) showed strong expression of a- smooth muscle actin.

结果：组织块贴壁法成功培养出SD大鼠胸主动脉平滑肌细胞，培养的细胞呈典型&峰-谷&样生长，HE染色细胞呈梭形，胞浆丰富，核大而圆或椭圆，免疫组化S-P法检测a-平滑肌肌动蛋白单克隆抗体呈强阳性表达。

TyPe II collagen induced arthritisln the rat ank1e joint andoVathumin as antigen induced arthritis WA in the rabbit knee joint wereestab1ish2 Qualitative evaluation of me in skin, muscle, synovium, cedilagearound joint and blood was performed by OMA3 The CIA rats were treated on day 7 after hind paw swelling and erythemaAnimals were injected intravenously with ase at a dose of 10mg/kg,tWenty minuots 1ater, one ankle of the rats random1y assigned was exPosedlaser irradiation at l00J/cm fOr l000 seconds, and another ankle wasM grouP wihout laser The other two groups is unmanipulatedcontrol group and untreated CIA group Bimaleolar ankle widthmeasuremellts were taken in all animals every tWo days using amicrometer The histopathology of the ank1e Joint was assessed at day 21after disease onset4 The pro1iferating cell nuclear antigen WCNA of CIA treated by PDT andthe HMME group without laser was doterdrined by immunohistochemiStry5 The AfA rabbits were treated on day 7 after knee swelling and erythemaThe theraPy invo1ved lntravenous injection of l0mg/kg HMME, fOl1owedby 20 minues period in dim light, and transdermal light treatment with\l00 J/cm2 fOr l000 seconds The inner sides of the treated Anees wereirradiated at first, and then the outer side did 24 hours later, the synovialtissue of the Anees joint were removed and in situ cel1 aPoptosis wasdetCCted With tednal deoxync1eotidyl transferase-mediated dUTP nickend labelingR6suIt8:l The pathologic changes of CIA and AIA include subsynovial inflammation,opovial hyPerplasia, pannus formation, cartilage and bone destructionresemble RA.2 The studies demonstrated that there are different uptake of HMME withinskin, muscle, synovium, cartilage and b1ood, and the synovium cou1draPidly uPtake more ase than skin and cartilage at the firSt 30 minuesaller intravenous injection of HMME3 The bimaleolar anke width had no different among PDT treated group,H group withollt 1aser and untreated CIA group But hlstologicalevaluation showed statiStical1y significallt reductions in synovialhyperplasia, pannus formation and cart1lage reosion, bone destruction andtotal score in PDT treated group4 Image analysis showed that the ratlo bforeen the areas of the coufltedobect to that of the entire area in PDTtreated grOup is lower than that in conirol group, but the integrated oPticaldensity had no different between the two groups5 Imape analysis showed that the ratio between the area of the countedobject to that of the e

治疗组在大鼠出现踝关节红肿后1周，炎症达到高峰时进行PDT治疗。随机治疗大鼠一侧的踝关节，另。2。一一侧作单纯HMME 对照。治疗方法是大鼠麻醉后尾静脉注入 HMME10ngkg，20分钟后踝关节照光，激光波长627.sum，功率密度 100mwcm'，照射时间1000秒，能量密度100）／。治疗后避光喂养72 小时。隔日一次测量大鼠的踝关节左右横径，治疗后两周取关节进行病理d 观察。 4。大鼠CIA模型用上述方法进行PDT治疗后，治疗组和单纯HMME 组用兔疫组化SP法检测石蜡切片的核增殖抗原。 5。兔AIA模型在关节炎出现第七天进行PDT治疗，随机治疗一侧膝关节，另一侧作自身对照。兔耳静脉注入I'arrainrelomg／Kg，20分钟后，膝关节用金蒸气激光照射，激光能量密度100）儿旷。24 ／J'时后取膝关节滑膜作病理检查，并用脱氧核昔酸末端转移酶介导的缺口末端标记法原位检测凋亡细胞。结果： 1。模型观察：CIA大鼠炎症高峰期滑膜下炎细胞浸润明显，滑膜细胞明显增殖，炎症达到高峰后二周，血管缀形成，并侵蚀和破坏软骨和骨， CIA模型病理改变与人类RA相似。兔AIA模型膝关节滑膜病理可见滑膜细胞增生，滑膜下炎细胞浸润，也与人类RA滑膜改变相似。 2。关节周围组织中光敏剂含量的测定结果表明，各组织对HMME 的吸收速度和吸收量不同，荧光值一时间曲线不同，滑膜组织比皮肤和软骨对 HMME的吸收多，在 2 0分钟时即有明显差异。 3.PDT对CIA模型的治疗结果表明：PDT治疗后关节炎组、单纯 HMME组和治疗组踝关节左右横径统计学检验差异没有显著性，但病理评分PDT治疗组滑膜增生、血管资形成及软骨破坏、骨破坏和总分比关节炎对照组和HMME对照组好，统计学检验差异有显著性。。3＿军医进修学院硕士学位论文中文摘要 4.PDT治疗组PCNA阳性细胞较对照组少，图像分析结果表明面密度（阳性染色的面积总和与统计视野面积的比值）治疗组小于对照组，统计学检验差异有显著性。。 5.PDT治疗组凋亡阳性细胞较对照组明显增多，图像分析结果单位视野内阳性细胞数和面密度PDT治疗组高于对照组，统计学检验差异有显著性。凋亡细胞核直径PDT治疗组较小，与对照组相比，统计学检验差异有显著性。结论：二。CIA、AIA的病理改变类似人类RA，可作为研究RA病因、发病机制、检查及治疗方法的模型。 2。各组织对HMME的吸收速度和吸收量不同，滑膜组织比皮。

AIM: To investigate the antineoplastic effect of pholiota nameko polysaccharides and the mechanism of inducing K562 cell apoptosis. METHODS: MTT assay was used to determine the inhibitory rate of PNP with different concentrations on the proliferation of K562 cells, and the cell growth curve was drawn.

采用MTT法检测PNP对K562细胞增殖的抑制作用；绘制生长曲线；Hochest 33258染色计算凋亡率；RTPCR法、Western Blot方法检测凋亡相关基因Caspase3的表达。

Open-field test,step-down avoidance test and immunohistochemstry staining were employedto study the effects of the extract of the fruit of Hyperieum perforatum on praxiology and cerebral pathology in the rat models.

制作大鼠慢性应激抑郁模型，应用敞箱法、跳台法、免疫组织化学染色研究贯叶连翘提取物对抑郁大鼠行为学、脑病理学的影响。

According to the result showed at 280nm and at 490nm,in the comparison of whether protein absorption top and sugar quantity top overlapped,glycoproteins would be detected preparatorily,and as a result,tubes in two distinct areas had glycoproteins by this method. Proteins were precipitated with trichloroaceticacid and with cold acetone,and glycoprotein was determined from SDS-gel.

再从各收集管的收集液中，用三氯乙酸沉淀蛋白法、冷丙酮沉淀蛋白法相结合浓缩、制备蛋白样品，进行SDS—PAGE，对SDS胶进行PAS糖链染色鉴定糖蛋白，并从茶树叶分级蛋白中准确地鉴定出两个区域的收集管中含有多种糖蛋白。

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。