siRNA沉默PKM2对胃癌BGC-823细胞增殖和迁移的抑制作用

在恶性肿瘤组织中,丙酮酸激酶（PK）的组织特异性同工酶（L型、R型、M1型PK）表达减少,而PKM2表达升高,在肿瘤细胞生长、增殖、迁移等方面发挥重要作用。目的：探讨siRNA沉默PKM2对胃癌BGC-823细胞增殖和迁移的抑制作用。方法：以蛋白质印迹法筛选出高表达PKM2的胃癌细胞株BGC-823。构建PKM2siRNA干扰质粒,稳定转染胃癌BGC-823细胞,同时以转染空质粒pU6作为阴性对照组。以荧光定量PCR和蛋白质印迹法在mRNA和蛋白质水平上验证干扰效果,以CCK8法和细胞迁移实验检测细胞增殖和迁移能力。结果：与阴性对照组相比,PKM2siRNA转染组PKM2mRNA和蛋白表达明显下降,细胞增殖能力和迁移能力明显下降（P〈0．05）。结论：PKM2在人胃癌的发展中发挥重要作用,并对胃癌BGC-823细胞增殖和迁移有一定的促进作用。     

RNAi沉默Sp3基因对肝癌HepG2细胞增殖的影响

目的:采用基因沉默和慢病毒转染技术,沉默人肝癌细胞HepG2中特异蛋白3(specificity protein 3,Sp3)的表达,观察沉默Sp3基因后的肝细胞的增殖能力变化.方法:采用Sp3-siRNA慢病毒转染人肝癌细胞HepG2,通过免疫印迹法和PCR技术检测Sp3的表达以验证转染效果,通过MTT检测细胞生长曲线和流式细胞技术测定细胞周期.结果:Western blot实验表明,实验组的Sp3表达量明显少于空白组和阴性对照组(0.37±0.08 vs 0.83±0.17,0.66±0.13,F=8.442,均P<0.05).RT-PCR也得到相同的结果(0.47±0.05 vs 0.74±0.08,0.70±0.16,F=7.322,均P<0.05).MTT实验结果显示,与空白对照和阴性对照组相比,实验组细胞在48、72和96h时的增殖明显受到抑制(0.28±0.18 vs 0.34±0.19,0.35±0.07,F=3.888;0.57±0.11 vs 0.84±0.05,0.74±0.08,F=12.721;0.72±18.1 vs 0.98±0.05,0.93±0.9,F=6.342,均P<0.05).流式细胞术结果显示实验组细胞主要分布在G1期.结论:RNAi沉默Sp3基因导致人肝癌HepG2细胞体外增殖能力下降.     

Increased parietal activity after training of interference control

Recent studies suggest that computerized cognitive training leads to improved performance in related but untrained tasks (i.e. transfer effects). However, most study designs prevent disentangling which of the task components are necessary for transfer. In the current study, we examined whether training on two variants of the adaptive dual n-back task would affect untrained task performance and the corresponding electrophysiological event-related potentials (ERPs). Forty three healthy young adults were trained for three weeks with a high or low interference training variant of the dual n-back task, or they were assigned to a passive control group. While n-back training with high interference led to partial improvements in the Attention Network Test (ANT), we did not find transfer to measures of working memory and fluid intelligence. ERP analysis in the n-back task and the ANT indicated overlapping processes in the P3 time range. Moreover, in the ANT, we detected increased parietal activity for the interference training group alone. In contrast, we did not find electrophysiological differences between the low interference training and the control group. These findings suggest that training on an interference control task leads to higher electrophysiological activity in the parietal cortex, which may be related to improvements in processing speed, attentional control, or both.     

RNA干扰沉默缺氧诱导因子-1α对人膀胱尿路上皮癌T24细胞增殖和凋亡的影响

目的 探究RNA干扰靶向沉默缺氧诱导因子-1α(HIF-1α)在人膀胱尿路上皮癌(BUC) T24细胞株中的表达对T24细胞增殖和凋亡的影响.方法 构建4个针对HIF-1α的小干扰RNA(siRNA)表达质粒PGC-siHIF-1α1-4,脂质体法稳定转染T24细胞.采用逆转录-聚合酶链反应(RT-PCR)和酶联免疫吸附试验(ELISA)检测转染细胞HIF-1α rnRNA和基因表达,噻唑蓝(MTT)法测定细胞增殖率,流式细胞仪(FCM)检测细胞周期和凋亡情况.结果 4个HIF-1α mRNA表达质粒均能不同程度的抑制转染细胞的HIF-1α膜mRNA和基因表达.其中以PGC-siHIF-1 α3的作用最大,效果最佳,该组G0/G1期阻滞率(61.8±1.5)％,细胞增殖受抑率41.7％,细胞凋亡率19.3％,与对照组有统计学差异(P＜0.01).结论 HIF-1α siRNA表达质粒可抑制膀胱癌细胞生长并促进其凋亡,有望成为提高膀胱癌分子靶向治疗的新位点.     

Deciphering IgM interference in IgG anti-HLA antibody detection with flow beads assays

In flow beads assays, the interference of IgM for IgG anti-HLA antibodies detection is not precisely understood. Using the screening flow beads assay for class I HLA antibodies, we analyzed the binding of two IgG mAbs, the anti-class I HLA W6/32 and an anti-beta-2-microglobulin, in the presence of an anti-beta-2-microglobulin IgM mAb. In neat serum, the IgM mAb impaired the detection of both IgG. In EDTA-treated serum, the interference was stronger for the anti-beta-2-microglobulin IgG than for W6/32, in agreement with the finding in surface plasmon resonance that this IgM competed with the anti-beta-2-microglobulin IgG but not with W6/32. The IgM interference was higher in neat than in EDTA-treated serum for both IgG mAbs. The IgM interference was also analyzed with class II single antigen flow beads and sera from two kidney recipients containing IgG and IgM donor specific antibodies. Anti-HLA IgG detection was partially corrected by EDTA, and restored by IgM inactivation with DTT, confirming the results observed with the mAbs. Therefore, three mechanisms can explain the IgM interference for IgG anti-HLA antibodies in flow beads assays: direct competition for antigen, steric hindrance and complement activation.     

一种难溶性原料药细菌内毒素检查方法的建立

摘 要 目的：建立原料药HSSYO 001 3S细菌内毒素检查方法。方法: 以二甲亚砜（DMSO）溶解HSSYO 001 3S,用适量细菌内毒素检查用水（BET水）稀释后,离心取上清液,按中国药典2015年版四部通则11431检查凝胶法,对HSSYO 001 3S进行细菌内毒素检查的方法学研究。结果:HSSYO 001 3S加助溶剂并以BET水稀释至1 mg·m^-1 ,经离心后取上清液稀释4倍及以上时对鲎试剂凝集反应无干扰作用。结论:HSSYO 001 3S可采用细菌内毒素检查法控制其质量。     

叔丁醇细菌内毒素检查方法研究

目的 为建立叔丁醇的细菌内毒素检查方法提供参考。方法 按《中国药典》2015年版（四部）要求验证叔丁醇预处理的可行性,并采用凝胶法考察样品对细菌内毒素检查的干扰情况。结果 样品水浴蒸发挥干后,加等体积BET水溶解残渣作为供试液,对试验无干扰作用。结论 初步建立叔丁醇的细菌内毒素检查方法。     

老年习惯性便秘的社区干预效果评估

目的探讨社区干预对老年习惯性便秘的效果。方法收集辖区内常驻老年习惯性便秘患者154例,在征得本人及家属同意,接受并积极配合本社区干预措施情况下,治疗3个月,并采用调查问卷的形式,对干预前、后患者便秘症状好转情况进行比对分析。结果135例老年习惯性便秘患者经社区干预性治疗3个月后,痊愈38例(28.15%)、好转83例(61.48%)、无效14例(10.37%),总有效率89.63%,且干预前后患者对便秘相关因素认知情况、心理干预情况均有显著差异,应用SPSS软件建立数据库并进行统计分析。以P<0.05差异有统计学意义。结论社区的正确干预,对老年习惯性便秘是行之有效的。为了早日缓解便秘患者的痛苦,提高生活质量,我们应大力推广。     

注射用尤瑞克林细菌内毒素检查方法的建立

目的:建立注射用尤瑞克林细菌内毒素的检查方法。方法:按照2010年版《中国药典》(二部)附录细菌内毒素检查法的要求,用2个厂家的鲎试剂对3个批号的注射用尤瑞克林进行干扰试验和细菌内毒素检查。结果:样品的浓度≤0.000 75 PNA/ml时对细菌内毒素检查无干扰。结论:细菌内毒素检查法可以用于注射用尤瑞克林细菌内毒素的限量检查。     

注射用兰索拉唑细菌内毒素检查法的建立

目的:建立注射用兰索拉唑的细菌内毒素检查法。方法:按照2010年版《中国药典》(二部)附录细菌内毒素检查法,采用2个厂家的鲎试剂对5个厂家的5批样品,通过预干扰和干扰试验确定样品最大无干扰质量浓度,并进行细菌内毒素检查。结果:将供试品溶液稀释到质量浓度为0.1 mg/ml时,不干扰细菌内毒素试验,其细菌内毒素限值为5 EU/mg。结论:采用细菌内毒素检查法检查注射用兰索拉唑中的细菌内毒素方法可行。