RNA干扰沉默HMGN5基因对肺癌H1299细胞增殖和周期的影响

目的:探讨RNA干扰技术沉默高迁移率族核小体结合域5(HMGN5)基因对肺癌H1299细胞增殖和细胞周期的影响,为肺癌的基因靶向治疗提供理论依据.方法:构建HMGN5特异性siRNA慢病毒载体,感染肺癌H1299细胞,设阴性对照组及干扰组,应用Real-time PCR和Western blotting方法分别从mRNA和蛋白质水平检测各组干扰质粒对HMGN5基因的干扰效果,MTT和BrdU法检测HMGN5 siRNA作用下的细胞增殖率,流式细胞仪检测细胞周期变化.结果:与阴性对照组比较,干扰组HMGN5的mRNA表达量下降了50.7%(P< 0.05),蛋白表达水平明显降低(P<0.05);MTT检测,干扰组细胞增殖水平明显低于阴性对照组;BrdU实验RNAi干扰组细胞增殖率(37.8%)明显低于对照组(55.0%)(P< 0.05);流式细胞仪检测细胞G1期细胞百分比(54.6%±0.9%)高于阴性对照组(46.5%±0.4%)(P< 0.05).结论:运用RNA干扰技术能够有效沉默H1299细胞的HMGN5基因,并抑制肺癌细胞的增殖能力,提示HMGN5在肺癌的发生发展中起重要作用,抑制HMGN5的表达可能成为一种治疗肺癌的方法.     

茶多酚与银杏叶提取物联用对人乳腺癌细胞增殖的影响

[目的]研究茶多酚与银杏叶提取物联合作用对人乳腺癌细胞 MDA-MB-231增殖的影响.[方法]采用MTT 法测定茶多酚、银杏叶提取物单独及联合应用对人乳腺癌细胞系MDA-MB-231的抑制作用;利用光镜观察茶多酚、银杏叶提取物对人乳腺癌系MDA-MB-231形态的影响.[结果]MTT 法显示银杏叶提取物与低浓度(10、20μg/ml)的茶多酚联合使用,对乳腺癌细胞抑制率明显高于单独用药组,但是联合用药组只有在一定的浓度范围内才会产生协同作用抑制乳腺癌细胞增殖;在细胞形态方面,与单独用药组、对照组相比,联合用药组乳腺癌细胞生长状况差,折光性弱,贴壁细胞皱缩,变圆.[结论]银杏叶提取物在低浓度下对人乳腺癌细胞系MDA-MB-231 生长无明显抑制作用,茶多酚和茶多酚联合银杏叶提取物均可抑制MDA-MB-231 细胞生长,联合用药组强于单独用药,在一定浓度范围内起协同作用.     

添加氧化铁对8YSZ导电性能的影响研究

用柠檬酸溶胶-凝胶法制备了添加氧化铁的8YSZ前驱体凝胶,凝胶在500℃煅烧后,压制成φ12mm×2mm圆片,在1 400℃烧结4h得到所需试样.研究了氧化铁的不同添加量对试样的烧结性能和电导率的影响.结果表明,添加氧化铁可提高试样的烧结性能,但使试样的电导率下降.     

热处理对Ti-8V-8Cr-2Zr-3A1合金丝材组织和性能的影响

分别选用不同的固溶温度、固溶冷却方式和时效保温时间对Ti-8V-8Cr-2Zr-3A1合金丝材进行热处理,研究了热处理制度对丝材室温拉伸性能及金相显微组织的影响。结果表明：固溶处理温度在740～820℃范围内,力学性能及显微组织对温度不敏感,固溶温度继续提高时,材料晶粒明显变大,强度降低,塑性下降;固溶温度为740℃,分别以水淬、空冷方式冷却,材料的组织、性能无明显变化,以炉冷方式冷却,强度提高,塑性下降。眼镜架用Ti-8V-8Cr-2Zr-3A1合金丝材较为理想的热处理工艺为740℃×10min／AC＋550℃×（4～6）h／AC,经该工艺处理后,材料的抗拉强度约1120MPa,延伸率大于12％,能够满足用户要求。     

小干扰RNA逆转三氧化二砷耐药的白血病K562/AS2细胞Topo Ⅱ基因表达的实验研究

目的 研究小干扰RNA片段(shRNA)对三氧化二砷(ATO)耐药的白血病细胞株K562/AS2细胞的Topo Ⅱα、TopoⅡβ基因表达及其功能的影响.方法 设计并合成针对Topo Ⅱα和TopoⅡβ基因序列的shRNA各3对,在脂质体的介导下转染K562/AS2细胞;用荧光实时定量聚合酶链反应(PCR)分析Topo Ⅱα、TopoⅡβmRNA的表达水平;流式细胞术检测Topo Ⅱα、TopoⅡβ蛋白表达.结果 针对Topo Ⅱα-shRNA、TopoⅡβ的shRNA作用于K562/AS2细胞24 h后,Topo ⅡαmRNA水平和蛋白水平最大下调为(78.22±0.01)%、(31.17±1.27)%(P<0.05),TopoⅡβmRNA水平和蛋白水平最大下调为(57.36 ±0.01)%、(23.98 ± 1.22)%(P<0.05).结论 转染24 h后针对TopoⅡ的shRNA可抑制对ATO耐药的白血病细胞株K562/AS2细胞TopoⅡ基因的表达.     

富血小板血浆对体外培养条件下人真皮成纤维细胞增殖能力的影响

目的:探讨富血小板血浆(PRP)对人真皮成纤维细胞(hDFbs)在体外培养条件下增殖能力的影响,探讨PRP促进皮肤、黏膜伤口愈合的机制.方法:PRP和hDFbs来源于健康成年人,两次离心法制备PRP,倒置相差显微镜观察0、12.5%、25.0%、50.0%和100.0%PRP浓度作用下 hDFbs的增殖;免疫细胞化学检测50.0%浓度不同剂量PRP作用下细胞血小板源性生长因子(PDGF)的表达;荧光染色技术观察PRP作用下hDFbs在纯钛材料表面的生长;流式细胞术检测PRP培养后不同时间hDFbs细胞周期;CCK-8法检测不同浓度PRP培养条件下细胞增殖活力.结果:倒置相差显微镜下见PRP各浓度组细胞数量均多于对照组,细胞数量增加、折光性增强;免疫细胞化学检测,30 μL PRP组PDGF表达量最高且细胞密度最大,但10 μL PRP组累积吸光度值(IOD)高于20 μL PRP组(836.27±21.15 vs 794.35±30.26,P<0.05);荧光染色技术观察,PRP组材料表面hDFbs细胞密集,数量较对照组高;细胞周期检测,PRP促进细胞进入S期进行DNA复制,PRP作用后第2天PRP组S期细胞百分比高于空白组(34.41% vs 22.00%,P<0.05),第8天PRP组G0/G1期细胞百分比高于空白组(95.07% vs 89.70%,P<0.05);CCK-8测定细胞增殖活性,100.0%PRP组吸光度A450值高于12.5%PRP组(34.41% vs 22.00%,P<0.05).结论:高浓度的PRP虽然表现较强的促细胞增殖作用,但并不存在浓度、剂量依赖性,适宜浓度的PRP可促进hDFbs的增殖.     

甲基化寡核苷酸灭活死亡相关蛋白激酶1基因对白血病K562细胞增殖的影响

目的 探讨互补甲基化寡核苷酸诱导灭活K562白血病细胞死亡相关蛋白激酶1基因(DAPK1)及对其增殖的影响.方法 应用Lipo2000将与DAPK1基因启动子序列互补的甲基化寡核苷酸转染进K562白血病细胞,分别应用甲基化特异性聚合酶链反应(MSP)和反转录PCR(RT-PCR)检测转染前后DAPK1基因启动子甲基化状态和mRNA表达改变.应用噻唑蓝(MTT)法检测转染前后细胞增殖变化.结果 正常组K562细胞的DAPK1基因启动子表现为未甲基化状态,可检测到相应mRNA表达;对照组寡核苷酸转染后,DAPK1基因启动子表现为未甲基化状态,mRNA表达和细胞增殖速度与正常组无明显差异;互补甲基化寡核苷酸转染后,DAPK1基因启动子呈甲基化状态,mRNA呈低表达状态,细胞增殖速度较正常组、甲基化对照寡核苷酸转染组显著增加.结论 互补甲基化寡核苷酸可诱导灭活K562白血病细胞DAPK1基因并抑制其mRNA表达,促进细胞增殖.     

加工工艺对TC8钛合金组织和拉伸性能的影响

测试了TC8合金在不同轧制温度和热处理工艺下的室温及500℃下的拉伸性能,观察了合金的显微组织。结果表明,合金的拉伸性能主要与轧制的变形量和热处理的冷却速度有关,空冷可以获得较佳的强度与塑性的结合;变形量越大,所获的合金室温拉伸性能越高,而变形量在70％以上可以获得较好的高温（500℃）拉伸性能。     

Ti8LC钛合金热处理工艺对硬度和组织的影响

钛合金Ti8LC是一种新型低成本近а型的钛合金,为了研究该合金热处理对其组织结构的影响规律,对其进行不同温度固溶处理,随后在不同温度进行时效热处理.用金相显微镜和扫描电镜观察了不同热处理工艺下的微观组织形貌,并通过能谱分析了不同工艺下元素的含量变化.采用洛氏硬度计测试了试样的平均硬度,分析了固溶温度和时效温度对其微观组织形态的变化影响,以及硬度大小、微观组织结构中相的形态和组织组成的相关性.结果表明:固溶处理温度对Ti8LC固溶后的显微组织及随后的时效组织都有极大的影响,随固溶温度的升高,由片状逐渐转为块状.经相同时效工艺处理的合金其显微组织受固溶后组织的遗传影响,合金的硬度随着固溶温度的升高.先降低后升高;通过微观组织和硬度的良好配合,可以确定Ti8LC合金固溶时效的最佳工艺.     

稀土氧化物对再生YG8硬质合金性能的影响

以锌熔法回收的WC-Co粉末为原料,采用常规硬质合金生产工艺制备添加不同含量Y₂O₃和Nd₂O₃的再生YG8硬质合金,并对其进行密度、硬度、抗弯强度以及断口形貌检测分析,探讨了稀土氧化物对硬质合金的强化机理。结果表明,添加Y₂O₃的硬质合金钴磁值升高,添加Nd₂O₃的钴磁值降低;稀土氧化物Y₂O₃和Nd₂O₃对硬质合金密度、硬度影响均不大,添加量为0.5%时,硬度达到最大,分别为89.5、89.3 HRA;抗弯强度随着稀土氧化物含量的增加明显提高,添加0.7%Y₂O₃和0.7%Nd₂O₃的硬质合金抗弯强度分别达2 200、2 102.8 MPa,比未添加稀土氧化物的再生YG8硬质合金提高了22.2%和16.8%;添加Y₂     

Coproduction of Regan isoenzyme and carcinoembryonic antigen in HCT-8 cells

Five groups of adult Ss were submitted to free-recall learning of 24 objects presented as written nouns for the control group and as pictures for experimental groups, as follows: schematic black and white, schematic colored, detailed black and white, and detailed colored drawings. Pictures were better recalled than nouns. Recall was significantly inferior for schematic black and white than for detailed colored drawings, with schematic colored and detailed black and white drawings at an intermediate level. These results support the so-called incidental-cues hypothesis according to which the best recall of pictures in comparison with that of nouns is partially the result of pictures having more distinctive cues likely to be encoded than do verbal items.     

Cell proliferation induced by 8-oxoguanosine and 8-methylguanosine, two adducts produced by free radical attack on ribonucleosides and RNA

The vowel "eh" was used to study auditory-nerve responses at high sound levels (60-110 dB). By changing the playback sampling rate of the stimulus, the second formant (F2) frequency was set at best frequency (BF) for fibers with BFs between 1 and 3 kHz. For vowel stimuli, auditory-nerve fibers tend to phase-lock to the formant component nearest the fiber's BF. The responses of fibers with BFs near F2 are captured by the F2 component, meaning that fibers respond as if the stimulus consisted only of the F2 component. These narrowband responses are seen up to levels of 80-100 dB, above which a response to F1 emerges. The F1 response grows, at the expense of the F2 response, and is dominant at the highest levels. The level at which the F1 response appears is BF dependent and is higher at lower BFs. This effect appears to be suppression of the F2 response by F1. At levels near 100 dB, a component 1/component 2 transition is observed. All components of the vowel undergo the transition simultaneously, as judged by the 180 degrees phase inversion that occurs at the C2 transition. Above the C2 threshold, a broadband response to many components of the vowel is observed. These results demonstrate that the neural representation of speech in normal ears is degraded at high sound levels, such as those used in hearing aids.     

Lectin-mediated bioadhesion: binding characteristics of plant lectins on the enterocyte-like cell lines Caco-2, HT-29 and HCT-8

Lipophilic esters of the naturally occurring polyamines putreanine and spermic acid were synthesized, characterized, and modified with nitric oxide to form the corresponding 1-substituted diazen-1-ium-1,2-diolates (previously known as NONOates). The resulting compounds were insoluble in water but were able to release nitric oxide (NO) when placed in phosphate buffered saline (PBS) at physiological pH. The two categories of compounds examined were putreanate esters of cholesterol or hexadecanol, and spermate diesters of cholesterol or hexadecanol. The putreanate NONOate esters of cholesterol and hexadecanol had NO release half-lives of 60 h and 81 h, respectively while the spermate NONOate diesters of cholesterol and hexadecanol and NO release half-lives of 23 days and 7.1 days, respectively. The presence of 5% Tween 20 increased the half-life of the cholesteryl putreanate NONOate to 104 h but only slightly increased the half-life of the hexadecyl putreanate NONOate to 89 h. The 1% presence of the pharmaceutical lung surfactant Survanta did not significantly alter the half-life of NO release for the cholesteryl putreanate NONOate or for the hexadecyl spermeate NONOate, but the surfactant did increase the amount of NO release by 46% and 67% respectively.     

Influence of metoclopramide on the pharmacokinetics of 8-methoxypsoralen

VISX 20/20 excimer laser was used to correct myopia from 1.00 to 16.00 diopters on 529 myopia eyes in 306 patients. All the patients were followed up for 3 to 12 months (8.1 +/- 3.1 months). At the 12th postoperative month, 0.5-3 grade corneal haze occurred in 62.1% of eyes in high myopia eyes, and only 8.8% in low and moderate myopia eyes (P < 0.01). The factors of formation of the corneal haze were the central corneal thickness, methods of removing epithelium and the number of laser pulses. The results suggest that the photorefractive keratectomy is regarded safe in correction of low and moderate myopia, but not in high myopia.     

Cell death and cell proliferation contribute to the enhanced growth of foci by fasting in rat medial colon

We have recently shown that fasting before initiation markedly stimulated the growth of aberrant crypt foci (ACF) induced by azoxymethane (AOM) in the rat medial colon. Here we investigated the mechanisms by which fasting enhanced the growth of ACF. Rats were exposed to 4 day-starvation, then they were given AOM (20 mg/kg) on the first day of refeeding. 4 day-fasting depressed cell proliferation as shown by the decreased mitotic index and enhanced cell death by apoptosis. On the first day of refeeding, apoptotic index remained higher than control values, while mitotic index markedly increased in the colonic epithelium of fasted/ refed rats. The administration of AOM induced an apoptotic wave, that was higher in controls, and a transient drop in the mitotic index that recovered quickly in the fasted/refed group. These data suggest that starvation-induced apoptosis represents the mitogenic stimulus to increase the rates of cell proliferation responsible for the enhanced growth of ACF in fasted/refed rats.     

Prevention by retinoids of azoxymethane-induced tumors and aberrant crypt foci and their modulation of cell proliferation in the colon of rats

Retinoids are proposed chemopreventive agents that inhibit cell proliferation and induce differentiation. Their ability to prevent azoxymethane (AOM)-induced aberrant crypt foci (ACF) and tumors and to modulate cell proliferation was investigated in the colon of male F344 rats. Thirteen retinoids were evaluated for prevention of ACF and two of them, 9-cis-retinoic acid (RA) and 4-(hydroxyphenyl)retinamide (4-HPR), were also evaluated for prevention of colon cancer. The retinoids were administered continuously in the diet starting 1 week prior to the first of two weekly 15 mg/kg i.p. injections of AOM and for a total of either 5 or 36 weeks in order to evaluate their effect on colonic ACF and tumors. At a concentration of 1 mmol/kg diet, 2-(carboxyphenyl)retinamide caused the greatest reduction (57.7%) in the yield of ACF. 9-cis-RA was toxic at 1 mmol/kg so that it was evaluated at 0.1 mmol/kg, resulting in a 41.6% reduction in ACF. The ability of the retinoids to reduce the proliferating cell nuclear antigen (PCNA) labeling index in ACF and in non-involved crypts correlated with their ability to prevent ACF. Both 9-cis-RA (0.1 and 0.2 mmol/kg diet) and 4-HPR (1 and 2 mmol/kg diet) were highly effective in decreasing the yield of AOM-induced colon tumors. In summary, retinoids were demonstrated to reduce cell proliferation and to prevent ACF and tumors in the colon, suggesting promise as preventive agents for colon cancer.     

Influence of spices on the bacterial (enzyme) activity in experimental colon cancer

Using degenerate PCR primers that target evolutionarily conserved sequences in pal genes, we show that in the gymnosperm, Pinus banksiana, phenylalanine ammonia-lyase (PAL) is encoded by a multigene family of at least eight to ten loci. Five classes of pal sequence were easily distinguished among 28 clones sequenced from the products of PCR amplification of haploid genomic DNA. The dominant sequence from each class was named, yielding pal1 to pal5 loci. These genes shared 68.8% to 94.0% nucleotide identity over the 366 bp region compared. All of pal1 to pal5 were expressed in cell suspension cultures treated with a fungal elicitor and all but pal3 were expressed in differentiating xylem tissue of a mature tree. Only pall was expressed in unelicited cell cultures. While these P. banksiana genes are quite divergent, they are still more similar to each other than to any angiosperm pal gene cloned to date. For its roles in development and defense, PAL production in P. banksiana is coordinated from a large, diverse multigene family. We discuss evidence suggesting that other pines have similar pal gene family structures.     

Perforin and Fas killing by CD8+ T cells limits their cytokine synthesis and proliferation

During an immune response, effector CD8+ T cells can kill infected cells by the perforin-dependent pathway. In comparison to CD4+ T cells, which are major sources of cytokines, normal CD8+ T cells produced less interleukin 2 and interferon gamma, and proliferated less vigorously after antigenic stimulation. Killing of target cells was a major cause of these reduced responses, since perforin-deficient CD8+ T cells showed substantially increased cytokine synthesis and proliferation. Cytotoxicity by the alternate Fas pathway also resulted in self-limitation of CD8+ T cell cytokine synthesis. This relationship between cytotoxicity and cytokine synthesis may regulate CD8+ T function in different phases of an immune response.     

Influence of destruction of retina-RPE complex on the proliferation of scleral chondrocytes in chicks

BACKGROUND: Restenosis remains the major limitation of coronary angioplasty. Coronary stents have reduced the incidence of restenosis in selected patients with relatively large vessels. No strategies to date have demonstrated a beneficial effect in vessels < 3.0 mm in diameter. We have shown in the MultiVitamins and Probucol (MVP) Trial that probucol, a potent antioxidant, reduces restenosis after balloon angioplasty. The purpose of this study was to determine whether the benefit of probucol therapy is maintained in the subgroup of patients with smaller coronary vessels. METHODS AND RESULTS: We studied a subgroup of 189 patients included in the MVP trial who underwent successful balloon angioplasty of at least one coronary segment with a reference diameter < 3.0 mm. One month before angioplasty, patients were randomly assigned to one of four treatments: placebo, probucol (500 mg), multivitamins (beta-carotene 30000 IU, vitamin C 500 mg, and vitamin E 700 IU), or probucol plus multivitamins twice daily. The treatment was maintained until follow-up angiography was performed at 6 months. The mean reference diameter of this study population was 2.49+/-0.34 mm. Lumen loss was 0.12+/-0.34 mm for probucol, 0.25+/-0.43 mm for the combined treatment, 0.35+/-0.56 mm for vitamins, and 0.38+/-0.51 mm for placebo (P=.005 for probucol). Restenosis rates per segment were 20.0% for probucol, 28.6% for the combined treatment, 45.1% for vitamins, and 37.3% for placebo (P=.006 for probucol). CONCLUSIONS: Probucol reduces lumen loss and restenosis rate after balloon angioplasty in small coronary arteries.     

Enhancement of experimental colon cancer by genistein

Several phytochemicals and micronutrients that are present in fruits and vegetables are known to exert cancer chemopreventive effects in several organs, including the colon. Among them, the soybean isoflavonoid genistein received much attention due to its potential anticarcinogenic, antiproliferative effects and its potential role in several signal transduction pathways. The present study was designed to investigate the effect of genistein on azoxymethane (AOM)-induced colon carcinogenesis and to study its modulatory role on the levels of activity of 8-isoprostane, cyclooxygenase (COX), and 15-hydroxyprostaglandin F2alpha dehydrogenase (15-PGDH) in the colonic mucosa and colon tumors of male F344 rats. At 5 weeks of age, groups of male F344 rats were fed control (AIN-76A) diet or a diet containing 250 ppm genistein. Beginning 2 weeks later, all animals except those in the vehicle-treated groups were given weekly s.c. injections of AOM (15 mg/kg body weight) for 2 successive weeks. All rats were continued on their respective dietary regimen for 52 weeks after AOM treatment and were then sacrificed. Colon tumors were evaluated histopathologically. Colonic mucosae and tumors were analyzed for COX, 15-PGDH, and 8-isoprostane levels. Administration of genistein significantly increased noninvasive and total adenocarcinoma multiplicity (P < 0.01) in the colon, compared to the control diet, but it had no effect on the colon adenocarcinoma incidence nor on the multiplicity of invasive adenocarcinoma (P > 0.05). Also, genistein significantly inhibited the 15-PGDH activity (>35%) and levels of 8-iosoprostane (50%) in colonic mucosa and in tumors. In contrast, genistein had no significant effect on the COX synthetic activity, as measured by the rate of formation of prostaglandins and thromboxane B2 from [14C]arachidonic acid. The results of this investigation emphasize that the biological effects of genistein may be organ specific, inhibiting cancer development in some sites yet showing no effect or an enhancing effect on the tumorigenesis at other sites, such as the colon. The inhibition of 8-isoprostane levels by genistein indicates its possible antioxidant potential, which is independent of the observed colon tumor enhancement, yet this agent may also possess several biological effects that overshadow its antioxidant potential. The exact mechanism(s) of colon tumor enhancement by genistein remain to be elucidated; it is likely that its colon tumor-enhancing effects may, at least in part, be related to inhibition of prostaglandin catabolic enzyme activities.