Comparative proteomic analysis of differentially expressed proteins between K562 and K562/ADM cells

Background Multidrug resistance to chemotherapeutic agents is an important clinical problem during the treatment of leukemia. The resistance process is multifactorial. To realize the total factors involved in multidrug resistance, we analyzed the differentially expressed proteins of K562 and K562/ADM cells and we investigated one of the up-regulated proteins （CRKL） using siRNA to determine its role in K562/ADM cells. Methods Altered protein expressions between K562/S （K562 ADM-sensitive cell line） and K562/ADM （K562 multidrug resistant cell line induced by adriamycin） were identified by 2D-DIGE coupled with mass spectrometry. Meanwhile, we confirmed the differential expression of CRKL and Stathmin in both K562 and K562/ADM cells by Western blot analysis. Furthermore, we used RNA interference to silence the CRKL gene expression. Results Among the 9 differentially expressed proteins, 3 were up-regulated in K562/ADM cells, while 6 were down-regulated in the K562/ADM cells compared with its parent cell line. The expression of CRKL was up-regulated significantly in K562/ADM cells, and it can be decreased by recombinant lentivirus. Moreover, the multidrug resistance of K562/ADM cells was efficiently reversed by silence of CRKL gene expression. Conclusions The data provided the differentially expressed proteins in K562 and its resistant cell line and highlights the power of 2D-DIGE for the discovery of resistance markers in cancer. We found CRKL may be a new protein involved in the multidrug resistanse of leukaemia cells.     

The protective effects of in vitro cultivated calculus bovis on the cerebral and myocardial cells in hypoxic mice

Summary The protective effects of in vitro cultivated calculus bovis (ICCB) on the cerebral and myocardial cells in hypoxic mice and the mechanism were examined. In one group, mice were intragastrically (i.g.) given ICCB for 15 days and then they were subjected to acute cerebral ischemia by decapitation, and then the panting time was recorded. In the other group, 12 min after exposure to hypoxia, mice was administered the ICCB i.g. for 5 days, and then the blood serum and tissues of brain, heart, liver were harvested and examined for SOD, GSH-px and T-AOC activity and content of MDA. The tissues of brain and heart were observed electron-microscopically for ultrastructural changes. The corpus striatum and hippocampus of brain were collected and examined for content of dopamine (DA) and norepinephrine (NE). The ultrastrural examination showed that the pathological change in brain and heart in the ICCB group was very slight, while abnormal changes in the control group were obviously more serious. ICCB significantly prolonged the panting time of the hypoxic mice (P<0.001), increased the activity of SOD, GSH-px, T-AOC in serum and tissues of brain, liver, heart and elevated the content of DA and NE. ICCB also pronouncedly reduced content of MDA in serum and tissues of brain, heart and liver. Significant differences in these parameters were noted between ICCB group and controls. It is concluded that ICCB can exert protective effect on the cells of brain and myocardium by enhancing the tolerance of the tissues to hypoxia and the body’s ability to remove free radicals and regulating the neurotransmitters.     

儿童亚急性坏死性淋巴结炎病因及临床特征

目的探讨亚急性坏死性淋巴结炎(SNL)的病因、实验室检查及临床特征,提高对儿童SNL的认识,减少误诊,合理治疗。方法结合文献资料,回顾分析13例SNL患儿的临床资料。结果本组13例患儿经淋巴结活检确诊为SNL,以学龄期儿童为主,中位年龄10.3岁,男女之比约为2.3∶1;临床特征表现为发热、浅表淋巴结肿大、白细胞及中性粒细胞减少(82%),可伴有肝或脾轻度肿大(30%);急性期多有病毒及支原体感染;免疫学检查:T细胞亚群测定显示CD4+/CD8+降低(63%);病理特点为淋巴结广泛的凝固性坏死伴组织细胞反应性增生;广谱抗生素治疗无效,复发1例,其余病儿预后良好。结论对不明原因发热伴淋巴结大的学龄患儿,除行相关实验室检查外,应早期行淋巴结活检以明确诊断。SNL可能与病毒感染或变态反应有关,存在细胞免疫功能紊乱,病程多呈自限性,糖皮质激素治疗敏感,亦有复发,应长期随访。     

Effects of sodium ferulate on the ultrarapid delayed rectifier K^＋ current in human atrial myocytes

Objective To study the effects of sodium ferulate on the ultrarapid delayed rectifier K current(Ikur) in human atrial myocytes.MethodsHuman atrial myocytes were isolated by enzyme dispersion method. Ikur in human atrial myocytes were recorded by using the whole cell patch clamp. The changes of Ikur were compared in the absence and the presence of sodium ferulate. ResultsThere was no effect of 0.4 g/L sodium ferulate on I-V relation of Ikur However, 0.4 g/L sodium ferulate inhibited Ikur to some degrees at each test pulse. The current densities of Ikur at 60 mV decreased from 4.997 ± 0.35 PA/PF to 3.331 ± 0.26 PA/PF(n = 6, P ＜ 0.05). The inhibitory effect was concentration-dependent. IC50 was(0.41 ± 0.03)g/L and the Hill coefficient was 0.95 ± 0.05. ConclusionSodium ferulate as a potassium channel blocker can inhibit Ikur in human atrial myocytes effectively.     

吡咯烷二硫氨基甲酸酯对犬体外循环中红细胞的保护作用

目的研究犬体外循环(CPB)中吡咯烷二硫氨基甲酸酯(PDTC)对红细胞的保护作用。方法将12只犬随机分为对照组(n=6)和PDTC组(n=6),建立CPB模型。PDTC组于CPB前静脉注射PDTC30mg/kg,对照组于CPB前静脉注射等量生理盐水。分别于CPB前、阻断30min、阻断60min、开放30min和开放60min时测定血浆白细胞介素(IL)-lβ、IL-8、游离血红蛋白(F-HB)、丙二醛(MDA)、红细胞三磷酸腺苷(E-ATP)和红细胞超氧化物歧化酶(E-SOD)水平。结果阻断后各时间点对照组的血浆IL-lβ和IL-8水平均显著高于CPB前(P<0·01),PDTC组开放后血浆IL-lβ和IL-8水平显著高于CPB前(P<0·05,P<0·01);两组血浆MDA和F-HB水平均显著高于CPB前(P<0·01);两组E-ATP和E-SOD水平均显著低于CPB前(P<0·01)。PDTC组CPB后E-ATP水平和E-SOD活性显著高于对照组(P<0·01),而血浆IL-lβ、IL-8、MDA含量和F-HB浓度均显著低于对照组(P<0·01)。结论PDTC通过减轻CPB期间脂质过氧化反应和炎性反应对红细胞起到一定的保护作用。     

腺病毒介导BCSC-1基因实验性治疗鼻咽癌

目的探讨抑癌基因BCSC-1对肿瘤的治疗作用。方法制备重组腺病毒Ad5-BCSC-1。用CellTiter 96 A-queous细胞增殖单一溶液检测试剂盒检测细胞增殖。肿瘤体内注射Ad5-BCSC-1,以注射Ad5-egfp或PBS作为对照,1周后重复注射,再2周后处死小鼠,称瘤重。结果Ad5-BCSC-1感染的CNE-2L2细胞的生长明显减慢,细胞在培养中聚集成大细胞团块。注射PBS、Ad5-egfp和Ad5-BCSC-1的小鼠CNE-2L2肿瘤的平均瘤重分别为(2·28±0·73)、(2·07±0·40)和(0·58±0·32)g。Ad5-BCSC-1组显著低于PBS和Ad5-egfp组(P<0·05)。结论瘤体内注射Ad5-BCSC-1能抑制CNE-2L2肿瘤的生长,提示BCSC-1基因治疗对BCSC-1基因缺失或表达低下的肿瘤可能有治疗作用。     

用SF—36量表评价慢性病患者的生命质量

目的 用SF－36量表分析慢性疾病对生命质量的影响。方法 在四川省某城乡用SP－36量表对438名慢性疾病患者进行调查,并按照ICD－8进行疾病分类,对不同系统疾病患者的生命质量进行分析。结果 不同系统疾病患者的生命质量在SF－36量表所测的不同领域均低于一般人群（≥40岁）,不同系统疾病患者在躯体活动性能、社会功能、活力和心理功能四个领域均有差别。影响生命质量的主要疾病有精神疾患、血液及造血器官疾病,传染病和寄生虫病。结论 慢性病降低了患者的生命质量。不同系统疾病对生命质量的影响程度不同,SF－36量表适用于评价慢性病患者的生命质量。在慢性疾病的预防控制中,生命质量应作为健康评价指标。     

血清及高密度蛋白对氧磷酶活性的分光光度测定法

目的 在现有的实验条件下,获得稳定可靠的血清及高密度脂蛋白（high density lipoprotein,HDL) 中对氧磷酶J（paraoxonase,PON）活性的测定方法。方法 以乙酸苯酯为底物,在对氧磷酶的作用下水解,生成产物苯酚,测定5分钟内反应体系中在270nm处苯酚特征性的紫外吸收,换算为酶促反应速度。结果 通过观测底物浓度、pH、激活剂Ca^2 、抑制剂EDTA等对酶促反应的影响,得出最适的测定条件;底物浓度为5mmol/L,最适pH为8．0,必需激活剂Ca^2 浓度为2mmol/L。结论 在该反应条件下,测定不同浓度HDL或血清中该酶的活性,重复性和稳定性均符合要求。     

四川省居民生命质量影响因素的研究

目的 找出影响生命质量的主要因素,为增进人群的生命质量提供参考依据。方法 用SF－36量表评价2249名四川居民的生命质量,同时调查可能影响生命质量的社会经济、文化因素、以及被调查对象的个人特征和慢性病情况。用t检验,方差分析和逐步回归对每一种影响因素分别分析,找出主要影响因素。结果 单因素分析时,是否患慢性病、年龄和性别、文化程度、婚姻状况、职业、经济收入、家庭人口及家庭代数、城乡分布、医疗费用负担形式等11种因素与生命质量的各领域或大多数领域均有关系。但进一步的多因素分析,在排除了各因素之间的相互影响后发现,对生命质量的主要影响因素是疾病、经济收入、城乡分布、年龄和文化程度,婚姻对个别领域有影响。结论 注意慢性病的预防与控制,积极发展经济,提高人民的收入水平和文化教育程度是提高人群生命质量的关键;分城乡和年龄制定SF－36的人群正常参数值很有必要,是否分性别制定尚需进一步研究。