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41.
目的:建立一种简单、稳定、可靠的PON1基因多态性分析法。方法:取外周静脉血100μl,用ROSE法提取基因组DNA,用两对引物:P192F 5′-TAT TGT TGC TGT GGG ACC TGA G-3′,P192R 5′-CAC GCT AAA CCC AAA TAC ATC TC-3′;P55F 5′-GAA GAG TGA TGT ATA GCC CCA G-3′,P55R 5′-TTT AAT CCA GAG CTA ATG AAA GCC-3分别进行PCR扩增,用限制性内切酶AlwI,NlaⅢ对两种PCR产物分别进行酶切,3%琼脂糖凝胶电泳。结果:扩增的两个PCR产物在192、55多态性位点大小分别为99bp、170bp。Alw I酶切后,凝胶电脉得到完全酶切(66bp,33bp),部分酶切(99bp、66bp、33bp),未被酶切(99bp)三种类型DNA片段(RR,QR,QQ基因型);NlaⅢ酶切后,凝胶电脉得到部分酶切(170bp,126bp,44bp),未被酶切(170bp)两种类型DNA片段(LM,LL基因型)。经双盲重复检测,结果一致。应用此方法对胃癌患者血样标本检测,发现其PON1基因多态性在192位点频率较高。结论:采用一步PCR-RFLP技术可以建立简单、稳定、可靠的PON1基因多态性分析法。 相似文献
42.
二甲基亚砜诱导人肝癌细胞BEL-7402凋亡的研究 总被引:2,自引:0,他引:2
目的 :研究二甲基亚砜 (DMSO)对人肝癌细胞凋亡的诱导作用。方法 :用不同浓度的DMSO处理体外培养的人肝癌细胞BEL 74 0 2 ,应用普通光镜、荧光显微镜、MTT分析方法和流式细胞技术 (FCM )检测肝癌细胞凋亡的形态学变化、细胞存活率、凋亡百分率和细胞周期分布的变化。结果 :DMSO诱导BEL 74 0 2细胞核DNA凝缩和核片段化 ,最后形成凋亡小体 ;随着DMSO浓度的增加和处理时间的延长 ,细胞存活率明显下降 ,其IC50 为 1.9% ;2 %的DMSO处理细胞 12h ,凋亡率达 17.2 1% ,同时S期细胞明显增加 ,G2 M期细胞明显下降。结论 :DMSO可诱导人肝癌细胞凋亡 ,并使细胞受阻于S期而进入凋亡程序。 相似文献
43.
大鼠脑组织单胺类递质及其代谢产物的检测方法研究 总被引:13,自引:0,他引:13
目的 :研究大鼠脑组织中单胺类递质及其代谢产物的高效液相反相离子对色谱测定法。方法 :采用LiChrosorbC18,10 μm色谱柱 ,流动相为甲醇 :水 (4 0 :60 ) ,含 0 .0 2 8g LEDTANa2 ,0 .15g LSDS ,0 .2ml LH2 SO4(pH 2 .5~ 3 ) ,荧光检测波长 :λEX=2 85 ,λEM=3 3 3。结果 :对 87只大鼠脑组织中 4种单胺类递质及其代谢产物的含量进行了同时测定 ,高香草酸 (HVA) 2 .5 0~ 40 .0 μg ml、去甲肾上腺素 (NE) 0 .0 1~ 0 .5 0 μg ml、多巴胺 (DA)0 .0 5~ 1.0 0 μg ml、5 羟色胺 (5 HT) 0 .0 2 5~ 0 .5 0 μg ml,峰面积与其含量呈良好的线性关系。 结论 :该法操作简便、快速、准确 ,为组织中单胺类递质及其代谢产物检测的一种理想方法 ,并适用于临床相关研究。 相似文献
44.
目的 探讨在多柔比星 (阿霉素 )肾病综合征 (NS)幼年大鼠肾损伤过程中核因子 (NF) κB和血管紧张素ATⅠ、ATⅡ的表达及其相关性。方法 4周龄雄性Wistar大鼠单侧肾切除加腹腔注射阿霉素造成NS模型 ,分别以免疫组织化学和原位杂交检测ATⅠ、ATⅡ和NF κB。结果 肾病组随着病变时间的延长 ,NF κB和ATⅠ、ATⅡ表达的强度和部位均呈增强趋势 ,治疗组在相同时间点则两者都有不同程度下调 (P <0 .0 5 )。结论 在阿霉素肾病损伤过程中NF κB和ATⅠ、ATⅡ起着介导作用。 相似文献
45.
Adrenergic signalling between rat taste receptor cells 总被引:2,自引:0,他引:2
Scott Herness† Fang-li Zhao Namik Kaya Shao-gang Lu Tiansheng Shen Xiao-Dong Sun‡ 《The Journal of physiology》2002,543(2):601-614
In taste buds, synaptic transmission is traditionally thought to occur from taste receptor cells to the afferent nerve. This communication reports the novel observation that taste receptor cells respond to adrenergic stimulation. Noradrenaline application inhibited outward potassium currents in a dose-dependent manner. This inhibition was mimicked by the β agonist isoproterenol and blocked by the β antagonist propranolol. The α agonists clonidine and phenylephrine both inhibited the potassium currents and elevated intracellular calcium levels. Inwardly rectifying potassium currents were unaffected by adrenergic stimulation. Experiments using the RT-PCR technique demonstrate that lingual epithelium expresses multiple α (α1a, α1b, α1c, α1d, α2a, α2b, α2c) and β (β1, β2) subtypes of adrenergic receptors, and immunocytochemistry localized noradrenaline to a subset of taste receptor cells. Collectively, these data imply strongly that adrenergic transmission within the taste bud may play a paracrine role in taste physiology. 相似文献
46.
Ming Zhao Cheng Simon C F Rawlinson Andrew A Pitsillides Gul Zaman Subburaman Mohan David J Baylink Lance E Lanyon 《Journal of bone and mineral research》2002,17(4):593-602
The mechanism by which mechanical strain and estrogen stimulate bone cell proliferation was investigated using monolayer cultures of human osteoblastic TE85 cells and female human primary (first-passage) osteoblasts (fHOBs). Both cell types showed small but statistically significant dose-dependent increases in [3H]thymidine incorporation in response to 17beta-estradiol and to a single 10-minute period of uniaxial cyclic strain (1 Hz). In both cell types, the peak response to 17beta-estradiol occurred at 10(-8) - 10(-7) M and the peak response to strain occurred at 3500 microstrain ((mu)epsilon). Both strain-related and 17beta-estradiol-related increases in [3H]thymidine incorporation were abolished by the estrogen receptor (ER) modulator ICI 182,780 (10-8 M). Tamoxifen (10(-9) - 10(-8) M) increased [3H]thymidine incorporation in both cell types but had no effect on their response to strain. In TE85 cells, tamoxifen reduced the increase in [3H]thymidine incorporation associated with 17beta-estradiol to that of tamoxifen alone but had no such effect in fHOBs. In TE85 cells, strain increased medium concentrations of insulin-like growth factor (IGF) II but not IGF-I, whereas 17beta-estradiol increased medium concentrations of IGF-I but not IGF-II. Neutralizing monoclonal antibody (MNAb) to IGF-I (3 microg/ml) blocked the effects of 17beta-estradiol and exogenous truncated IGF-I (tIGF-I; 50 ng/ml) but not those of strain or tIGF-II (50 ng/ml). Neutralizing antibody to IGF-II (3 microg/ml) blocked the effects of strain and tIGF-II but not those of 17beta-estradiol or tIGF-I. MAb aIR-3 (100 ng/ml) to the IGF-I receptor blocked the effects on [3H]thymidine incorporation of strain, tIGF-II, 17beta-estradiol, and tIGF-I. HOBs and TE85 cells, act similarly to rat primary osteoblasts and ROS 17/2.8 cells in their dose-related proliferative responses to strain and 17beta-estradiol, both of which can be blocked by the ER modulator ICI 182,780. In TE85 cells (as in rat primaries and ROS 17/2.8 cells), the response to 17beta-estradiol is mediated by IGF-I, and the response to strain is mediated by IGF-II. Human cells differ from rat cells in that tamoxifen does not block their response to strain and reduces the response to 17beta-estradiol in TE85s but not primaries. In both human cell types (unlike rat cells) the effects of strain and IGF-II as well as estradiol and IGF-I can be blocked at the IGF-I receptor. 相似文献
47.
48.
[目的]探讨在关节镜下膝关节后侧腔室联合手术入路的重要性和可操作性。[方法]经过前内侧、前外侧和股骨髁切迹以及后内侧、后外侧和后纵隔内切口联合入路分别入镜、入器械,进行膝关节后侧腔室的探查和手术操作。[结果]216例(239膝)应用联合入路探查和治疗,其中5例膝因关节僵硬操作失败;175例膝用于治疗后侧腔室疾病,膝关节后侧腔室手术视野显著改善,探查和手术操作完善,均达到手术目的。1例膝内侧隐神经不全损伤,没有腘后神经、腓总神经、腘后血管、交叉韧带等重要组织损伤。[结论]膝关节后侧腔室病变较多,是检查和治疗的重要部位,并非“技术盲区”。这种联合手术入路,手术风险低,具备可操作性,可以提高手术效率和质量,可作为膝关节镜下常规手术入路。 相似文献
49.
Su-Ping Sun Ye-Ning Jin Hong-Peng Yang Yi Wei Zhao Dong 《World journal of gastroenterology : WJG》2007,13(39)
AIM: To evaluate the relationship between changes in serum transforming growth factor β1 (TGFβ1) level and curative effect of radiotherapy (RT) in patients with esophageal carcinoma.METHODS: Ninety patients with histologically confirmed esophageal carcinoma were enrolled. Serum samples for TGFβ1 analysis were obtained before and at the end of RT. An enzyme-linked immunosorbent assay was used to measure serum TGFβ1 level. Multivariate analysis was performed to investigate the relationship between disease status and changes in serum TGFβ1 level.RESULTS: Serum TGFβ1 level in patients with esophageal carcinoma before RT was significantly higher than that in healthy controls (P < 0.001). At the end of RT, serum TGFβ1 level was decreased in 67.82% (59/87) of the patients. The overall survival rate at 1,3 and 5 years was 48.28% (42/87), 19.54% (17/87)and 12.64% (11/87), respectively. Main causes of death were local failure and regional lymph node metastasis.In patients whose serum TGFβ1 level decreased after RT,the survival rate at 1, 3 and 5 years was 61.02% (36/59),28.81% (17/59) and 18.64% (11/59), respectively. The survival rate at 1 year was 17.86% (5/28) in patients whose serum TGFβ1 level increased after RT, and all died within 18 mo (P < 0.01).CONCLUSION: Serum TGFβ1 level may be a useful marker for monitoring disease status after RT in patients with esophageal carcinoma. 相似文献
50.
本文报告用间接免疫荧光法检测特异性IgG抗体对流行性出血热的诊断意义。53例不同病期的出血热患者的血清,其间接荧光抗体的阳性率为49/53(占92.45%)。有77.36%的患者血清,检出IgG抗体的时间是在发病后的1~2周。血清中IgG的滴度与疾病的严重性无密切相关。另25例非EHF患者,其血清特异性IgG抗体均为阴性。可见检测特异性IgG抗体对EHF的诊断颇为有用。 相似文献