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81.
82.
高滴度单克隆抗体杂交瘤细胞系获得途径的探讨 总被引:1,自引:0,他引:1
为探索获得能分泌较高价单克隆抗体杂交瘤细胞系的途径,比较了用不同免疫方法得到的小鼠脾细胞制备杂交瘤,对其产生有效瘤和高价抗体分泌瘤的影响。用不使小鼠致病的病毒(如NDV)或巳灭活的病毒作为抗原时,以较高浓度加佐剂、长间隔(30天)、多次免疫效果较好。用能使小鼠感染的病毒作为免疫原时,以活病毒的亚致死剂量感染小鼠,取其脾细胞制备杂交瘤,效果最理想。有效杂交瘤产率高,分泌高效价抗体的杂交瘤细胞系也多。探讨了从免疫方法着手,定向研制高价单克隆抗体杂交瘤细胞系的可行性。 相似文献
83.
Tian W Han X Yan M Xu Y Duggineni S Lin N Luo G Li YM Han X Huang Z An J 《Biochemistry》2012,51(2):724-731
Overactivation or overexpression of β-catenin in the Wnt (wingless) signaling pathway plays an important role in tumorigenesis. Interaction of β-catenin with T-cell factor (Tcf) DNA binding proteins is a key step in the activation of the proliferative genes in response to upstream signals of this Wnt/β-catenin pathway. Recently, we identified a new small molecule inhibitor, named BC21 (C(32)H(36)Cl(2)Cu(2)N(2)O(2)), which effectively inhibits the binding of β-catenin with Tcf4-derived peptide and suppresses β-catenin/Tcf4 driven reporter gene activity. This inhibitor decreases the viability of β-catenin overexpressing HCT116 colon cancer cells that harbor the β-catenin mutation, and more significantly, it inhibits the clonogenic activity of these cells. Down-regulation of c-Myc and cyclin D1 expression, the two important effectors of the Wnt/β-catenin signaling, is confirmed by treating HCT116 cells with BC21. This compound represents a new and modifiable potential anticancer candidate that targets β-catenin/Tcf-4 interaction. 相似文献
84.
Herbaceous model species, especially Arabidopsis has provided a wealth of information about the genes involved in floral induction and development of inflorescences and flowers. While the genus Populus is an important model system for the molecular biology of woody plant. These two genuses differ in many ways. This study was designed to improve understanding of flower development in poplar at a system level, as its regulatory pathway to a large extent remains poorly known, owing to the presently limited mutant pool. To address this issue, a poplar GeneChip was employed to detect genes expressed during the whole floral developmental process. Using the expressed floral genes, a systematic gene network was constructed with the aid of functional association with Arabidopsis. The results suggested that autonomous, gibberellin, vernalization, photoperiod, ethylene, brassinosteroid, stress-induced and floral suppression pathways are involved in poplar flowering. Modularity analysis revealed several pathways in common with Arabidopsis, such as autonomous, gibberellin, vernalization and photoperiod pathways. In addition, brassinosteroid, stress-induced and floral suppression pathways were implicated as additional novel pathways. Notably, a difference in vernalization between Arabidopsis and poplar was revealed. Autonomous, gibberellin, vernalization, photoperiod, ethylene, brassinosteroid, stress-induced and floral suppression pathways integrated into a systematic gene network in floral development of poplar. Compared to Arabidopsis, brassinosteroid, stress-induced and floral suppression pathways are additional in poplar, and FLC is absent in vernalization pathway in poplar. Preliminary conclusions drawn here provide a basis for both identification of key genes and elucidation of molecular mechanisms involved in poplar floral development. 相似文献
85.
17个新的C2H2型锌指基因片段的分离与克隆 总被引:4,自引:0,他引:4
按照C2H2型锌指基因保守结构域的DNA序列设计一对简并引物,以人基因组DNA为模板进行PCR同源扩增,将由此获得的锌指基因片段为探针,从人胎肾、骨骼肌、骨骼组织的cDNA分子库中筛选到22个C2H2型锌指蛋白cDNA片段,经国际NCBI数据库查询检索,其中17个为新的锌指基因片段。对从胎肾cDNA分子库中分离到的K3-4和K5-12克隆进行了表达谱分析,发现K3-4在肾脏中的表达量明显高于其他几 相似文献
86.
VPAC2在CHO细胞的表达及鉴定 总被引:1,自引:0,他引:1
PAC2是垂体腺苷酸环化酶激活多肽(Pituitary adenylate cyclase activating polypeptide,PACAP)和血管活性肠肽(vasoactive intestinal peptide,VIP)的共同受体,介导多种重要生物学功能。为获得稳定特异表达VPAC2的中国仓鼠卵巢(Chinesehamsterovary,CHO)细胞,将pcDNA-VPAC2表达载体转染CHO细胞,G418筛选转染阳性克隆,PACAP38标准品诱导阳性克隆细胞的胞内cAMP生成,筛选出对PACAP38最为敏感的阳性单克隆细胞株(VPAC2-CHO),运用RT-PCR、Westernblot和免疫荧光法检测VPAC2受体表达情况,利用VPAC2受体特异激动剂通过竞争性结合试验和促进胞内第二信使cAMP生成的活性检测实验证实,VPAC2-CHO特异表达有功能的VPAC2。Scatchard作图分析显示VPAC2-CHO的VPAC2受体密度为(1.1±0.2)pmol/mg膜蛋白,PACAP38与VPAC2的解离常数Kd值为(0.55±0.10)nmol/L。特异表达VPAC2受体细胞系的构建为深入研究该受体理化性质、生物学功能以及筛选、开发VPAC2受体新型特异激动剂和拮抗剂等研究奠定了基础。 相似文献
87.
Jitai Zhang Juan Bai Qian Zhou Yuxin Hu Qian Wang Lanting Yang Huamin Chen Hui An Chuanzan Zhou Yongyu Wang Xiufang Chen Ming Li 《Cell death & disease》2022,13(5)
The activation of pancreatic stellate cells (PSCs) is the key mechanism of pancreatic fibrosis, which can lead to β-cell failure. Oxidative stress is an important risk factor for PSC activation. There is no direct evidence proving if administration of glutathione can inhibit fibrosis and β-cell failure. To explore the role of glutathione in pancreatic fibrosis and β-cell failure induced by hyperglycaemia, we established a rat model of pancreatic fibrosis and β-cell failure. The model was founded through long-term oscillating glucose (LOsG) intake and the setup of a sham group and a glutathione intervention group. In vitro, rat PSCs were treated with low glucose, high glucose, or high glucose plus glutathione to explore the mechanism of high glucose-induced PSC activation and the downstream effects of glutathione. Compared with sham rats, LOsG-treated rats had higher reactive oxygen species (ROS) levels in peripheral leukocytes and pancreatic tissue while TGFβ signalling was upregulated. In addition, as the number of PSCs and pancreatic fibrosis increased, β-cell function was significantly impaired. Glutathione evidently inhibited the upregulation of TGFβ signalling and several unfavourable outcomes caused by LOsG. In vitro treatment of high glucose for 72 h resulted in higher ROS accumulation and potentiated TGFβ pathway activation in PSCs. PSCs showed myofibroblast phenotype transformation with upregulation of α-SMA expression and increased cell proliferation and migration. Treatment with either glutathione or TGFβ pathway inhibitors alleviated these changes. Together, our findings suggest that glutathione can inhibit PSC activation-induced pancreatic fibrosis via blocking ROS/TGFβ/SMAD signalling in vivo and in vitro.Subject terms: Pre-diabetes, Pre-diabetes 相似文献
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89.
目的 探讨支气管镜肺泡灌洗联合抗生素对小儿重症肺炎的疗效及对痰细菌清除率和其他指标的影响。 方法 选取我科收治的重症肺炎患儿126例,分为研究组及对照组,各63例。其中研究组在常规治疗基础上联合支气管镜肺泡灌洗及局部应用抗生素治疗,对照组采用常规治疗,比较2组患者的临床治疗效果。 结果 治疗后,研究组的临床有效率为90.5%,高于对照组的76.2%(χ2=4.629,P=0.031);治疗后研究组患者的痰液中病菌清除率为77.7%,显著高于对照组的57.2%(χ2=6.110,P=0.013);治疗后研究组患者的相关炎性指标(高敏C反应蛋白、肿瘤坏死因子α及白介素 6)均较对照组显著降低(t=3.522、4.912、4.183,P=0.033、0.032、0.033);治疗后研究组患者的相关血气指标(动脉血氧分压及氧合指数)均较对照组显著升高,而二氧化碳分压较对照组显著降低(t=3.612、3.312、6.162,P=0.039、0.040、0.028)。 结论 BAL联合抗生素对小儿重症肺炎的疗效更为显著,值得临床推广应用。 相似文献
90.