XIAP表达水平对5-Fu诱导肝癌细胞凋亡的影响

目的研究XIAP mRNA及蛋白表达水平对不同剂量5-氟尿嘧啶（5-fluorouracil,5-Fu）诱导人肝癌细胞HepG2凋亡的影响。方法脂质体介导含XIAP基因全长的质粒pef-XIAP转染人肝癌细胞HepG2,采用逆转录聚合酶链反应（RT—PCR）及western blotting检测XIAP基因和蛋白表达水平。通过不同剂量5-Fu诱导细胞凋亡,使用CCK-8试剂通过比色法分析检测5-Fu对转染pef-XIAP的HepG2细胞生长活性的抑制作用。结果各组HepG2细胞在不同剂量的5-Fu诱导下发生凋亡、与普通HepG2细胞组比较．转染XIAP基因的HepG2细胞组的凋亡率显著降低。结论XIAP基因在人肝癌细胞HepG2中的高表达可以明显降低化疗药物5-Fu诱导的细胞凋亡,其可能在肝癌化疗耐药中起一定作用。     

Survivin蛋白和AIF在胃癌组织中的表达

目的探讨Survivin和AIF在胃癌组织中的表达及对胃癌诊断和预后评估的价值。方法应用免疫组化方法检测我院56例胃癌手术标本的Survivin和AIF的表达情况,并对这些患者进行随访。结果Survivin和AIF在胃癌细胞中表达阳性率分别为71.43%(40/56)和66.07%(37/56),Survivin在癌旁组织和正常胃组织中无表达,AIF在癌旁组织和正常胃组织中低表达;Survivin和AIF在胃癌中表达无显著相关性;Survivin及AIF的阳性和阴性组1、3、5年累积生存率均有统计学差异。结论Survivin和AIF在胃癌组织中的阳性表达率明显高于癌旁组织和正常胃组织,可以作为判断胃癌患者预后的指标,但二者的表达无明显相关性。     

诱导血红素氧合酶-1及雷帕霉素对PDGF诱导的大鼠平滑肌细胞表型转化的抑制作用

目的观察血红素氧合酶-1(HO-1)与雷帕霉素对血小板源性生长因子-BB(PDGF-BB)诱导的大鼠血管平滑肌细胞(VSMCs)表型转化的作用。方法原代大鼠VSMCs给予PDGF-BB诱导表型转化。同时给予不同剂量的血红素氧合酶诱导剂氯血红素诱导HO-1的表达,和雷帕霉素共培养24h。结果PDGF-BB20nmol/ml作用原代VSMCs24h后,SM-α-actin与PCNA表达减弱,PCNA表达增强。氯血红素诱导HO-1后可以抑制PDGF-BB诱导的VSMCs表型改变,并且随剂量的增加,抑制强度增加。较低剂量的雷帕霉素可以抑制PDGF诱导的VSMCs表型转化。结论PDGF-BB亚型可以促进原代培养的大鼠VSMCs由收缩表型向合成表型转化,给予氯血红素诱导HO-1表达可以对抗PDGF-BB诱导的大鼠VSMCs表型转化,并呈剂量相关。     

电针足三里对脓毒症大鼠促炎症因子所致心肌损伤的保护作用

目的研究电针足三里穴(ST36)对脓毒症大鼠促炎症因子所致心肌损伤的保护作用。方法雄性SD大鼠32只,采用盲肠结扎穿孔术(CLP)制备大鼠脓毒症模型,随机分为电针(Electro-Acupuncture,EA)足三里组(EA组)、CLP+假电针(ShameEA)组(SEA组)、迷走神经切断(Vagotomy,VA)+CLP组(VA组)和迷切后CLP再电针组(VA/EA组)4组,每组8只。EA组持续针刺双侧足三里穴1h(2mA,2-100Hz);SEA组采用相同频率和强度刺激非经非穴(足三里穴外侧旁开0.5cm)。VA组先切断腹腔迷走神经再施行CLP术。各组大鼠于CLP术后6h,取血检测血浆肌酸激酶同工酶(CK-MB)活性;然后处死动物,取心肌组织,测定肿瘤坏死因子-α(TNF-α)水平、一氧化氮(NO)含量、随过氧化物酶(MPO)活性以及组织含水率。结果CLP术后6h,EA组心肌组织促炎症因子TNF-α、NO、MPO水平、CK-MB活性以及组织含水率均显著低于其余3组(P<0.05);VA/EA组和VA组TNF-α、NO、MPO水平、CK-MB活性及组织含水率均高于SEA组(P<0.05);VA/EA组与VA...     

人白细胞介素12双亚基共表达pVAX1-IRES-hIL12载体的构建与表达

目的构建下游可以共表达人白细胞介素12(hIL12)双亚基的双顺反子真核表达载体pVAX1-IRES-hIL12。方法通过搭桥PCR获得人白细胞介素12P35及P40双亚基的融合基因P35-F2A-P40,插入DNA疫苗载体pVAX1-IRES的下游,瞬时转染293-T细胞,ELISA检测融合基因的表达。结果酶切鉴定和序列分析表明融合基因与设计完全一致,融合基因在体外细胞培养液检测中获得分泌表达。结论该载体的成功构建可以为肿瘤基因疫苗研制提供免疫增效载体。     

早期经鼻气管插管对高血压脑出血患者微创治疗后肺部感染的影响

目的研究早期经鼻气管插管对高血压脑出血患者微创治疗术后肺部感染发生率、ICU治疗时间的影响。方法选取58例2007年9月-2009年5月间于我院急诊科就诊并拟行微创治疗的高血压幕上型脑出血患者。根据GCS评分分层后进行随机分组,处理组于就诊时予以早期经鼻气管插管经人工鼻吸氧,对照组予以鼻导管或面罩吸氧,观察术后72h肺炎并发症发生率及ICU治疗时间,研究数据选取Fisher确切概率法及成组t检验方法进行统计分析。结果不管在GCS评分13～15分或6～12分的高血压脑出血患者中,早期插管组与对照组在术后肺炎发生率方面的差异无统计学意义(GCS13～15:21.4%vs35.7%,P=0.6776;GCS6～12:60%vs80%,P=0.2678),但在ICU治疗时间方面早期插管组较对照组明显缩短,两者间差异具有显著统计学意义(GCS13～15:4.9dvs6.9d;GCS6～12:8.2dvs10.3d,P<0.01)。结论对于高血压脑出血拟行微创治疗的患者行早期经鼻气管插管可明显减轻术后肺部并发症,减少ICU治疗时间,减轻患者经济负担,促进康复。     

选择性肝动脉栓塞治疗多囊肝的相关研究

【摘要】 传统治疗症状性多囊肝的方法有外科开窗、经腹腔镜开窗、部分性肝切除、肝脏移植、经皮穿刺抽吸及注入硬化剂治疗等，均存在一定缺陷。近年发现，在常染色体显性遗传性多囊肾病患者中，肝脏的囊肿由发育良好的肝动脉分支参与供血，门静脉不参与供血，前者被认为是囊肿液体的“来源”，因此用经导管动脉栓塞术闭塞供应囊肿的肝动脉分支后，可阻断液体来源、使囊肿缩小甚至消失，从而缓解患者的症状。

     

ADEMO/D: An adaptive differential evolution for protein structure prediction problem

Protein Structure Prediction (PSP) is the process of determining three-dimensional structures of proteins based on their sequence of amino acids. PSP is of great importance to medicine and biotechnology, e.g., to novel enzymes and drugs design, and one of the most challenging problems in bioinformatics and theoretical chemistry. This paper models PSP as a multi-objective optimization problem and adopts ADEMO/D (Adaptive Differential Evolution for Multi-objective Problems based on Decomposition) on its optimizer platform. ADEMO/D has been previously applied to multi-objective optimization with a lot of success. It incorporates concepts of problem decomposition and mechanisms of mutation strategies adaptation. Decomposition-based multi-objective optimization tends to be more efficient than other techniques in complex problems. Adaptation is particularly important in bioinformatics because it can release practitioners, with a great expertise focused on the application, from tuning optimization algorithm’s parameters. ADEMO/D for PSP needs a decision maker and this work tests four different methods. Experiments consider off-lattice models and ab initio approaches for six real proteins. Results point ADEMO/D as a competitive approach for total energy and conformation similarity metrics. This work contributes to different areas ranging from evolutionary multi-objective optimization to bioinformatics as it extends the application universe of adaptive problem decomposition-based algorithms, which despite the success in various areas are practically unexplored in the PSP context.     

Reduced Aggregation and Cytotoxicity of Amyloid Peptides by Graphene Oxide/Gold Nanocomposites Prepared by Pulsed Laser Ablation in Water

A novel and convenient method to synthesize the nanocomposites combining graphene oxides (GO) with gold nanoparticles (AuNPs) is reported and their applications to modulate amyloid peptide aggregation are demonstrated. The nanocomposites produced by pulsed laser ablation (PLA) in water show good biocompatibility and solubility. The reduced aggregation of amyloid peptides by the nanocomposites is confirmed by Thioflavin T fluorescence and atomic force microscopy. The cell viability experiments reveals that the presence of the nanocomposites can significantly reduce the cytotoxicity of the amyloid peptides. Furthermore, the depolymerization of peptide fibrils and inhibition of their cellular cytotoxicity by GO/AuNPs is also observed. These observations suggest that the nanocomposites combining GO and AuNPs have a great potential for designing new therapeutic agents and are promising for future treatment of amyloid‐related diseases.     

液相色谱-串联质谱法测定人血浆中银杏内酯B的含量

建立测定人血浆中银杏内酯B的含量的LC-MS/MS方法。取正常人血浆50μL,加入2.5mol/L盐酸溶液10μL酸化,再加入内标2μg/mL银杏内酯C20μL,以1mL乙醚萃取后,取提取液500μL挥干溶剂,然后用30%的乙腈500μL溶解,取10μL进行LC-MS/MS测定。LC条件:色谱柱为ZorbaxC18柱(4.5*150mm,5μm);流动相:乙腈-水梯度洗脱;流速:0.6mL/min,柱温:35℃。质谱条件:ESI电离源;负离子模式;用于定量分析的离子对分别为m/z423.1→m/z367.2(银杏内酯B)和m/z439.1→383.2(内标:银杏内酯C)。该方法血浆中银杏内酯B的最低定量限为5ng,线性范围为5ng/mL~0.5μg/mL。本方法操作简便,灵敏度高,结果准确可靠,可用于该药物的含量测定,同时也可为临床药代动力学研究提供参考。