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Get high purity DCs by Cultured plastic-adherent monocytes isolated from healthy human peripheral blood with GM-CSF and IL-4 for 7 days. To observe the morphology of DCs by inverted phase contrast microscope ,electron microscope and laser confocal microscope. Analyse phenotype of DCs with flow cytometry. Investigate the endocytosis ability of DCs as a group by Horseradish peroxidase endocytosis assay. To appraise allogeneic mixed lymphocytes reaction of DCs by MTT reduction assay. Analyse the levels of IL-12 and TNF in liquids of cultured medium by ELISA and MTT reduction assay respectively. Soluble antigens of HCCs was obtained by 3 freeze-and-thaw cycles. Biological characteristics of HC soluble antigens pulsed DCs were monitored by flow cytometry. According to MTT reduction assay estimated the cell proliferation of self lymphocytes activated by HC antigens pulsed DCs. Get high purity BCG HSP 70 protein by SDS-PAGE electrophoresis and determined its biological activity with ELISA. Analyse phenotype of antigen pulsed DCs primed by BCG HSP70 with flow cytometry. By MTT reduction assay estimated the cell proliferation of self lymphocytes and the MLR of DC based vaccine. Analyse expression of HLA-DR molecule on surface of HCC lines. The IFN-γ mRNA in lymphocytes after actived by DC vaccine and the Fas-L expression on DC and DC vaccine primed lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes and nonspecific inhibition of liquids in culture medium against HCC lines were also tested. Detect expression of hAFP on four HCC lines with Cell-ELISA. Induce apoptosis of HCCs with actinomycin-D. Interaction of DCs and apoptotic cells was observed under transmission electron microscope. Growth inhibition test of DC against HCC lines was also performed. Establish the nude mouse model bearing human HC xenografts and indentify the characteristic of tumour by histochemistry and immunohistochemistry techniques. Prevent and treat transplanted human HC on nude mouse with Freezing and anabiotic HC specific lymphocytes.

用GM-CSF和IL-4从健康人外周血诱导DC;分别用倒置相差显微镜、电子显微镜及激光共聚焦显微镜观察DC形态;流式细胞术检测DC表型;HRP吞噬实验测定DC的群体内吞能力;MTT法检测同种异体混合淋巴细胞反应;ELISA法和MTT法分别测定DC培养上清液中IL-12和TNF水平;冻融法制备肝癌细胞可溶性抗原;流式细胞术检测负载肝癌可溶性抗原后DC的生物学特性;MTT法检测DC负载肝癌抗原后对自身淋巴细胞增殖的影响;SDS-PAGE制备电泳纯化BCG HSP70并鉴定纯度,ELISA测定活性;流式细胞术检测负载抗原DC经BCGHSP 70活化后的表型;MTT法检测肝癌DC疫苗对自身淋巴细胞增殖的影响和混合淋巴细胞反应;流式细胞术检测肝癌细胞表面HLA-DR表达;MTT法检测肝癌DC疫苗对自身淋巴细胞的活化;原位杂交法检测肝癌DC疫苗活化后的淋巴细胞IFN-γmRNA表达;流式细胞术检测DC和肝癌DC疫苗活化后淋巴细胞表面Fas-L;MTT法分别检测肝癌DC疫苗活化的淋巴细胞和其培养上清对肝癌细胞的特异性杀伤和非特异性抑制作用;Cell-ELISA检测人肝癌细胞hAFP表达;MTT法检测负载AFP表位肽和凋亡肝癌细胞DC对自身淋巴细胞增殖的影响;ELISA法和MTT法分别测定活化后淋巴细胞培养上清中TNF和IL-12水平;肝癌细胞凋亡的诱导和检测;DC吞噬凋亡肝癌细胞后的电子显微镜观察;DC对肝癌细胞的生长抑制试验;人肝癌裸鼠皮下移植瘤动物模型的建立及其组织学和免疫组织化学鉴定;DC及肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤;冻存和复苏后的肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤。

The stable clones are further identified by RT-PCR and Western blot; 6 MTT assay is used to investigate the effect of ZNRD1 on the cell growth of cells (AGS, SGC7901, MKN28, NIH3T3, GES-1); 7 Soft agar assay is used to investigate the effect of ZNRD1 on the clonality of cells (AGS, MKN28); 8 Nude mice assay is used to investigate the effect of ZNRD1 on the cell growth of gastric cancer cells (AGS, MKN28); 9 Flow cytometry is used to investigate the effect of ZNRD1 on the cell cycle distribution of cells (AGS, MKN28, NIH3T3, GES-1); 10 Flow cytometry is used to investigate the effect of ZNRD1 on the cell apoptosis of cells (AGS, MKN28, NIH3T3); 11 MTT assay is used to investigate the effect of ZNRD1 on the drug sensitivity of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR) in vitro; 12 SRCA is used to investigate the effect of ZNRD1 on the drug sensitivity of gastric cancer cells (SGC7901, SGC7901/VCR) in vivo; 13 Flow cytometry is used to investigate the effect of ZNRD1 on adriamycin accumulation of cancer cells (SGC7901, SGC7901/VCR, HL-60, HL-60/VCR); 14 Transmission electron microscope is used to investigate the effect of ZNRD1 on the sensitivity of SGC7901 cells towards drug-induced apoptosis; 15 Flow cytometry and DNA ladder assay are used to investigate the effect of ZNRD1 on the sensitivity of cells (SGC7901, SGC7901/VCR, HL-60/VCR) towards drug-induced apoptosis; 16 Microarray is used to investigate the profiling of ZNRD1-responsive genes in gastric cancer cells (AGS, MKN28, SGC7901, SGC7901/VCR); 17 RT-PCR and Western blot are used to identify the results of microarray; 18 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of cyclin D1; 19 Reporter gene assay is used to investigate the effect of ZNRD1 on the transcriptional activity of MDR1; 20 Kinase assay is used to investigate the effect of ZNRD1 on the activity of cyclin E-CDK2 kinase; 21 The antisensenucleic acids of p21 is used to inhibit the expression of p21, and flow cytometry is used to investigate the effect of p21 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 22 The antisensenucleic acids of p27 is used to inhibit the expression of p27, and flow cytometry is used to investigate the effect of p27 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 23 Liposome is used to up-regulate the expression of Skp2, and flow cytometry is used to investigate the effect of Skp2 on ZNRD1-induced cell cycle arrest in gastric cancer cells; 24 Western blot is used to investigate the effect of ZNRD1 on the stability of Skp2 and p27 in gastric cancer cells; 25 MVD assay is used to investigate the effect of ZNRD1 on the angiopoietic activity of gastric cancer cells; 26 ELISA is used to investigate the effect of ZNRD1 on the expression of VEGF165 in gastric cancer cells; 27 The roles of DARPP-32 in MDR of gastric cancer cells are investigated using gene transfection, MTT assay, SRCA, flow cytometry and DNA ladder assay.

应用杂交瘤技术制备ZNRD1的首个单克隆抗体;2)利用RT-PCR、Western blot和免疫组化检测ZNRD1在胃癌组织、胃炎组织、正常胃上皮组织、胃癌细胞和正常胃组织上皮细胞中的表达;3)构建ZNRD1的小干扰RNA载体,并测序鉴定;4)利用脂质体将ZNRD1的真核表达载体及其空载体转染胃癌细胞(AGS、SGC7901、MKN28)和小鼠成纤维细胞(NIH3T3),G418筛选后进行鉴定;5)利用脂质体将ZNRD1的小干扰RNA载体及其空载体转染药敏胃癌细胞(SGC7901)、正常胃组织上皮细胞(GES-1)、对长春新碱耐药的胃癌细胞(SGC7901/VCR)、药敏白血病细胞(HL-60)、对长春新碱耐药的白血病细胞(HL-60/VCR),G418筛选后进行鉴定;6)利用MTT实验检测ZNRD1高/低表达对细胞(AGS、SGC7901、MKN28、NIH3T3、GES-1)生长的影响;7)通过软琼脂克隆形成实验检测上调ZNRD1对AGS、MKN28细胞克隆形成能力的影响;8)通过裸鼠成瘤实验检测上调ZNRD1对AGS、MKN28细胞体内成瘤性的影响;9)通过流式细胞仪分析ZNRD1高/低表达对细胞(AGS、MKN28、NIH3T3、GES-1)的细胞周期的影响;10)通过流式细胞仪分析上调ZNRD1对细胞(AGS、MKN28、NIH3T3)的凋亡的影响;11)通过MTT实验检测ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR、HL-60、HL-60/VCR)体外药物敏感性的影响;12)通过肾包膜下移植法检测ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR)体内药物敏感性的影响;13)通过流式细胞仪分析ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR、HL-60、HL-60/VCR)内阿霉素蓄积和泵出的影响;14)通过透射电镜检测上调ZNRD1对SGC7901细胞凋亡敏感性的影响;15)通过流式细胞仪和DNA梯度试验检测ZNRD1高/低表达对细胞(SGC7901、SGC7901/VCR、HL-60)凋亡敏感性的影响;16)通过基因芯片检测ZNRD1高/低表达对胃癌细胞内基因表达谱的影响;17)利用RT-PCR、Western blot对基因芯片的结果进行鉴定;18)利用报告基因实验检测ZNRD1对cyclin D1的启动子活性的调节作用;19)利用报告基因实验检测ZNRD1高/低表达对MDR1的启动子活性的调节作用;20)利用激酶试验检测ZNRD1对cyclin E-CDK2 激酶活力的影响;21)利用反义核酸技术抑制p21的表达;通过流式细胞仪检测抑制p21对ZNRD1介导的细胞周期阻滞的影响;22)利用反义核酸技术抑制p27的表达;通过流式细胞仪检测抑制p27对ZNRD1介导的细胞周期阻滞的影响;23)利用脂质体转染法上调Skp2的表达;通过流式细胞仪检测上调Skp2对ZNRD1介导的细胞周期阻滞的影响;24)利用Western blot检测ZNRD1对p27和Skp2的蛋白稳定性的影响;25)利用微血管密度实验检测ZNRD1对AGS、MKN28细胞裸鼠移植瘤微血管形成的影响;26)利用ELISA检测ZNRD1对AGS、MKN28细胞培养上清和移植瘤匀浆中VEGF165含量的影响;27)利用脂质体转染法、MTT实验、肾包膜下移植法、流式细胞仪和DNA梯度试验检测新耐药相关分子DARPP-32对细胞(SGC7901、SGC7901/VCR、对阿霉素耐药的胃癌细胞SGC7901/ADR)多药耐药表型的影响;利用脂质体转染法和MTT实验检测下调ZNRD1对DARPP-32介导的胃癌多药耐药的调控作用。

objective to study the efficacy of the tr/patoc 1 slide agglutination assay for vaccination new pentavalent whole cell vaccine for leptospirosis.methods using tr/patoc 1 slide agglutination assay to detect the searial of serum specimens of 100 healthy people after vaccination for leptospirosis and comparing with the microsopic agglutination test.results both basic immunizaton and 20 days after the enhanced immunization,their positive seroconversion rates were higher than 90% in tr/patoc 1 slide agglutination assay,they were not signficantly different between slide agglutination assay and mat,the antibody titer was greatly lower in 90 days after basic immunization.conclusion the tr/patoc 1 slide agglutionation assay was one of useful assays in evaluation the lately immune efficency of leptospira vaccination.

目的 采用tr/patoc 1玻片凝集试验评价钩端螺旋体疫苗免疫效果及应有价值。方法采用tr/patoc 1玻片凝集试验检测100名健康人钩端螺旋体疫苗免疫后的血清抗体变化,并与显微镜凝集试验比较。结果无论是基础免疫还是加强免疫20?d后,玻凝试验抗体阳性率高达90%以上,与标准的显微镜凝集试验比较差异无统计学意义,玻片凝集试验检测的抗体持续时间较短,90?d后阳性率已大幅度降低,表明sat检测的抗体是早期抗体。结论 tr/patoc 1玻片凝集试验操作简易,可以作为基层卫生机构监测钩端螺旋体疫苗近期免疫效果的方法之一。

Since that tumor metastasis is accompanied with proteolytic degradation of the extracellular matrix and changed cell motility, MMP-2, MMP-9 and u-PA activity were also studied via gelatin zymography and casein zymography assay to show that they all were inhibited by Terminalia catappa leaves.

在 gelatin zymography 与 casein zymography assay 中也发现到榄仁树的叶萃取物可以抑制 SCC-4 细胞中 MMP-2、MMP-9 及 u-PA 的表现。

On the basis of purification and immunity of vitellin, antibodies of vitellin were obtained and used to measure the concentration of vitellin in embryo in different stages of embryonic development by enzyme-linked immunosorbent assay.

在采用纯化的卵黄磷蛋白免疫兔子而获得其抗体的基础上,运用酶联免疫吸附检测法(enZyme一linked ilnlnunosorbent assay,ELISA)分析测定了发育过程中胚胎内卵黄磷蛋白的含量。

In patients with acute PE, if IV UFH is chosen, we recommend that after an initial IV bolus (80 U/kg or 5,000 U), it is administered by continuous infusion (initially at dose of 18 U/kg/h or 1,300 U/h) with dose adjustment to achieve and maintain an APTT prolongation that corresponds to plasma heparin levels of 0.3 to 0.7 IU/mL anti-Xa activity by the amidolytic assay rather than administration as IV boluses throughout treatment, or administration without coagulation monitoring (Grade 1C).

急性PE患者,如果选择IV UFH,我们推荐初始快速注射80U/kg或者5000U,继之以18 U/kg/h 或 1,300 U/h持续静滴。根据APTT延长,或者监测血浆肝素水平在0.3'0.7IU/ml, amidolytic assay 分析抗Xa因子活性来调整UFH剂量,而不能持续静脉注射治疗,或者不监测凝血功能(1C)。

Additionally, MCF-7 cells were incubated in L-arginine-free medium with 10% dialyzed FBS for 1-7 days to measure their cell viability by using MTT assay.

另外再以含有10%透析过FBS的L-arginine缺乏培养基培养MCF-7细胞1-7天后,以MTT assay测定其细胞存活率。

This nonmutagenic amino compound 4 (via the investigation into Salmonella/mammalian microsome assay) was also employed as diazo moiety coupling with acet-J-acid to form a diazo acid dye 5. The new dye 5 can color wool at 2% or silk at 3% o. w. f. into black one. The investigation of the uptake of 5 and its color appearance showed that it was assigned to an acid bluish black dye and its blackness was more excellent than that of Acid Black 10B.

经过Salmonella/mammalian microsome assay测试,4表现为非诱变性,用4与乙醯J-酸作用得到新型双偶氮酸性染料(5)。5对羊毛和丝绸分别在2%与3%色度下浸染都得到黑色染样,考察了5的上染曲线,并对染样进行了测色研究,发现该染料属於酸性蓝黑染料,其黑度优於目前市场上流行的酸性黑10B。

Can reverse JA-induced migration inhibition JA-treated cells also showed significantly higher adhesion to laminin In conclusion these results indicate a novel anti-tumor role of JA via reducing reorganization of cytoskeleton increasing integrin 3 expression and adhesion ability to inhibit tumor cells migration Furthermore immunofluorescence staining of phalloidin/actin indicated that JA-treated cells exhibited cytoskeleton rearrangement and significant decreases in the number and length of filopodia Activity of Rho-family small GTPases was detected by a GST-pull down assay and showed that GTP-loaded Cdc42 and Rac were decreased in JA-treated cells In the future we hope these findings will be helpful to further understand the mechanisms underlying tumor progression in oral cancer and subsequently improve the cancer prevention and treatment

以抗体阻断integrin ?3可以有效使OC2恢复被JA所抑制的迁移能力。细胞贴附实验指出JA能提高OC-2在laminin环境下的贴附能力。当我们进一步以免疫萤光染色观察细胞,发现在JA处理下的OC-2,其外形、细胞骨架和细胞边缘的filopodia的密度和长度明显减少,利用GST-pull down assay侦测到Cdc42-GTP和Rac-GTP的表现量降低。推论爵床素A可能透过刺激口腔癌细胞大量表现integrin ?3,使细胞贴附能力增加或是透过改变细胞骨架来降低细胞迁移能力。希望本研究对於JA所做的抗癌及抗转移效应评估能提供口腔癌治疗在研发上一些帮助。

Although NS4B seems to increase the severity of cell apoptosis, only limited effect was detected by nucleosome ELISA assay and flow cytometry.

不过在nucleosome ELISA assay和flow cytometry的分析上,NS4B对细胞凋亡的影响则不明显。

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