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超免疫法

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Methods: Separate and abstract the effective components of Huangqi (Omni-Saponin、total-flavone) and Danshen (Danshen-ketone、 Danshen-Quinone). Applying L-Arginine methyl ester to pregnant rats by I. P to make Nitrogen Oxide Synthesis blocking models (uterus-placenta-fetus lacking blood and oxygen model or pregnant hypertension syndrome model). On day 12, 18 of pregnancy paunched the rats and took out the fetus and placenta/uterus. Measured the basal BP during pregnant period and BP、protein、basal activity each day. Observed placenta/deciduas ultramicroscopic structure by electron microscope and took count of languishment cells. Measured the expressive intensity of bcl-2、Bax of placental nutrient cells by SABC method.

一氧化氮合成酶阻滞剂L-精氨酸甲酯妊娠大鼠腹腔给药,制作一氧化氮合成阻滞模型(子宫-胎盘-胎儿缺血缺氧或妊娠高血压模型),妊娠第12、18天剖腹取胎、胎盘/子宫,妊娠期测基础血压,每日测血压、蛋白质量、基本生理活动;电镜观察胎盘/蜕膜超微形态结构,并进行细胞凋亡计数;免疫组化检测胎盘滋养细胞bcl-2、Bax表达强度;原位杂交法检测胎盘滋养细胞、血管内皮细胞VEGF、eNOS、iNOS、MCP-1、MMP9(基质金属蛋白酶9)mRNA表达。

Methods: Twenty-four Kkay mice were divided into three groups(n=8):diabetes group,methionine-diet group,folic acid and vitamin B12 treatment group. C57Bl/6 mice were used as controls(C57). Four months later,the mice were sacrificed and their serum levels of Hcy,folic acid and vitamin B12 were measured by fluorescence polarization immunoassay. Pathological changes in kidneys were examined with the periodic acid schiff method with periodic acid-silver methenamine staining under electron microscope. Expression of MMP-9 protein or mRNA was detected by RT-PCR with immunohistochemical staining. Results: Different morphological changes of DN were found in Kkay mice.

Kkay糖尿病鼠随机分3组:糖尿病组、高蛋氨酸组和叶酸、维生素B12组,C57BL/6鼠为对照(C57),以相应饲料喂养4个月后取材,测定血糖、血同型半胱氨酸、叶酸和维生素B12,以PAS、六氨银染色法光镜下及超微电镜观察各组DN病变,比较各组肾小球硬化指数;以免疫组化和半定量图像分析肾小球MMP-9蛋白表达,并以RT-PCR检测肾脏MMP-9 mRNA水平。

Western blot was used to measure HSP70 of ischemia and non-ischemia myocardium and the expression of HSP70 was analyzed semiquantitatively.

采用蛋白质免疫印迹法检测各组缺血区和非缺血区心肌组织HSP70含量,并检测血清一氧化氯、丙二醛含量及总超氧化物歧化酶、肌酸激酶活性变化。

E and Sirus red in supersaturated picric acid solution;the amount of HA in serum was measured with the method of radiative immunity;the quantitative expression of fibrosis factor—TGF、PDGF was measured with the method of immunity histochemistry.Results:Myocardial collagen in model group increased obviously compared with normal group from the results of electron microscope、H.E、Sirus red in supersaturated picric acid solution.

腹腔注射柯萨奇B3病毒建立病毒性心肌炎慢性期心肌纤维化小鼠模型,随机分为模型对照组、通心络低剂量组、中剂量组、高剂量组,并设正常对照组,用药四周后处死动物通过透射电镜观察心肌细胞超微结构;HE染色、苦味酸天狼星红染色观察心肌胶原数量;放免法测定血清透明质酸的含量;免疫组化染色观察致纤维化细胞因子—转化生长因子和血小板衍生生长因子的定量表达。

Methods:Establish the model of myocardial fibrosis in chronic stage of viral myocarditis by injecting CVB3 into Balb/c rats,divide these rats into model group、Tong xin luo low dose group、Tong xin luo medium dose group、Tong xin luo high dose group and set up normal group,after four weeks of giving medicine execute the animals and observe the ultrastructure of myocardial cell with tramission electron microscope,the quantity of myocardial collagen was observed by staining with H.E and Sirus red in supersaturated picric acid solution;the amount of HA in serum was measured with the method of radiative immunity;the quantitative expression of fibrosis factor—TGF、PDGF was measured with the method of immunity histochemistry.

腹腔注射柯萨奇B3病毒建立病毒性心肌炎慢性期心肌纤维化小鼠模型,随机分为模型对照组、通心络低剂量组、中剂量组、高剂量组,并设正常对照组,用药四周后处死动物通过透射电镜观察心肌细胞超微结构;HE染色、苦味酸天狼星红染色观察心肌胶原数量;放免法测定血清透明质酸的含量;免疫组化染色观察致纤维化细胞因子—转化生长因子和血小板衍生生长因子的定量表达。

Gene bcl-2 and bax expression in tissue sections were assayed with immunohistochemistry supersensitive kit with a streptavidin-biotinperoxidase complex method.

应用链霉菌抗生物素蛋白-生物素-过氧化物酶复合物法免疫组化染色超敏试剂盒检测血管壁bcl-2、bax基因表达情况。

HMGB1, IL-6 levels in plasma and ultrafiltrate were measured by enzyme linked immunosorbent assay.

采用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)测定血浆和超滤液HMGB1、IL-6水平。

Methods :we used step increased load running training way to establish a aerobic and fatigue training model of rat, then applied respectively HE staining、Masson staining and immunohistochemisty SABC method to observe the change of AVN tissue structure and Cx43 and Cx45 expression of Koch triangle; and used electron microscope to observe the ultramicroscopic structure change of AVN parenchymal cell of Koch triangle.

采用跑台训练方式,建立大鼠有氧训练和疲劳训练模型,分别采用普通HE染、Masson染色及免疫组化SABC法,观察Koch三角区AVN组织结构变化及其Cx43和Cx45的表达;采用电镜观察Koch三角区AVN实质细胞的超微结构变化。

The treatment group was given Qizhengxiaotong Emplastrum(0.75 g/kg),and the model group and blank group were given normal saline solution(0.5 mL/each)externally for 5 days on 10th day after the establishment of ...

末次给药后测量各组小鼠的痛行为,包括机械性痛觉超敏和热刺激痛觉过敏;将小鼠股骨进行X线摄片,影像学评估骨破坏;用免疫组化法检测各组局灶皮肤组织肿瘤坏死因子α、内皮素-1(ET-1)、白介素-1β(IL-1β)水平及脊髓后角P物质受体、c-fos及GFAP阳性反应神经元的表达。

Methods Wistar rats of 6 months old were randomly divided into normal control groupand IR group. Each group had eight rats. Euglycemic insulin clamp technique was used to determine IR and glycose infusion rate was used to IR. Cardiocyte apoptosis was evaluated by TUNEL. Heart weight and body weight were measured to calculate HW/BW. Fine structure of heart and ultramicrostructure of cardiocyte were observed.

6月龄Wistar大鼠,分为正常对照组和IR组,正常血糖胰岛素钳夹技术测定各组大鼠IR,葡萄糖输注速率表示IR情况;测量大鼠心脏重量和体重并计算心脏重量与体重之比;原位末端酶标记法检测各组大鼠心肌细胞凋亡情况;观察心脏微细结构和心肌细胞超微结构;免疫组化检测心脏间质Ⅰ、Ⅲ型胶原水平。

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