红树莓果中鞣花酸提取物的抑菌活性初步研究

测定分析了红树莓果中鞣花酸提取物对常见微生物的抑菌活性. 结果 表明,鞣花酸提取物对大肠杆菌、沙门氏菌、枯草杆菌等受试细菌具有较好的抑制作用.鞣花酸提取物对受试霉菌米曲霉、黑曲霉、以及受试酵母菌抑制作用较弱.     

Curcumin通过Wnt信号通路抑制乳腺癌细胞MCF-7增殖的研究

目的 观察姜黄素(Curcumin)对乳腺癌细胞MCF-7增殖的影响,以及对细胞内Wnt信号通路的影响,探索Curcumin可能存在的抑制乳腺癌细胞增殖的分子机制.方法 体外培养人乳腺癌细胞MCF-7,并用不同浓度的Curcumin作用不同的时间.用MTT检测Curcumin对MCF-7细胞生长情况的影响;流式细胞仪观察经Curcumin作用后细胞周期的改变;RT-PCR和Westernblot分别检测细胞内β -catenin和下游靶基因CyclinD1的mRNA和蛋白水平的表达.结果 MTT结果显示Curcumin可以抑制MCF-7的增殖,并具有剂量-时间依赖性.在浓度为20 μmol·L-1时,对细胞生长的抑制作用最为明显.流式细胞仪观察细胞周期的结果提示,Curcumin能够阻止MCF-7细胞由G1期进入S期,提高Go/G1期细胞的百分比.RT-PCR和Western blot结果显示,Curcumin显著降低了细胞内β-catenin和CyclinD1的mRNA和蛋白水平的表达,且呈剂量-时间依赖性.结论 Curcumin能够抑制MCF-7细胞胞浆内β -catenin蛋白进入胞核,阻断Wnt信号转导通路.进而抑制下游靶基因CyclinD1的表达,阻止MCF-7由G1期进入S期,有效抑制了MCF-7细胞的增殖.     

阿卡波糖对胰α-淀粉酶的抑制动力学研究

为探究以马铃薯淀粉为底物时阿卡波糖对胰α-淀粉酶(来自猪胰)的抑制动力学,采用3,5-二硝基水杨酸显色法及双倒数作图法(Lineweaver-Burk法),体外模拟人体生理环境,建立胰α-淀粉酶最佳反应体系。结果显示,阿卡波糖在体外对胰α-淀粉酶活性具有明显的抑制作用,且作用效果随阿卡波糖浓度升高而增强,其半抑制浓度IC_(50)=5.975×10~(-6)mol/L,即3.854×10~(-3)mg/mL。通过双倒数作图法得知,胰α-淀粉酶的Km=12.309 mmol/L,阿卡波糖对胰α-淀粉酶的抑制类型为非竞争性与竞争性抑制混合型。本研究证实了以马铃薯淀粉为底物测定阿卡波糖对胰α-淀粉酶的抑制作用更为灵敏,为其它抑制剂以马铃薯淀粉为底物时抑制胰α-淀粉酶活性的研究提供了比较依据;且证实了阿卡波糖对胰α-淀粉酶有较强的抑制作用,可为临床治疗2型糖尿病时减少阿卡波糖的副作用提供一定的参考。     

克氏螯虾壳聚糖的荧光标记及其抗菌活性的研究

目的建立克氏螯虾壳聚糖的荧光标记方法并研究其抗菌活性。方法利用荧光物质5（6）-羧-四甲基罗丹明-5-马来酰亚胺按不同比例标记克氏螯虾壳聚糖,检测标记后克氏螯虾壳聚糖对大肠埃希菌及金黄色葡萄球菌的抗菌效果,并用激光共聚焦显微镜观察壳聚糖对试验菌的作用靶位。结果克氏螯虾壳聚糖与荧光物质（v／v）9：1混合时,所得的荧光标记克氏螯虾壳聚糖对两种试验菌的抗菌活性保留50％。荧光标记克氏螯虾壳聚糖已进入细菌内部。结论控制荧光标记克氏螯虾壳聚糖抗菌活性基团的比例,不影响其抗菌的活性,可作为一种荧光探针,用于克氏螯虾壳聚糖抗菌作用靶位的研究。     

高速动车组S38C车轴疲劳强度及剩余寿命评价

本文通过实验获得S38C车轴的残余应力场、基本力学性能和断裂性能,采用表面单位压力法及二次迭代,在实物车轴中重建出径向梯度分布的压缩残余应力,基于实测载荷谱进行含缺陷S38C车轴的剩余寿命预测.分析结果表明:残余压应力对裂纹扩展有显著的抑制作用,深度小于4 mm的初始裂纹不扩展;经典Paris方程和NASGRO方程估算寿命分别约为28.5万公里和89.3万公里.当深度大于7 mm时,裂纹扩展速率迅速增大,因此,把7 mm作为S38C车轴损伤容限分析止裂判据比较合理.研究方法和结果对于含缺陷表面强化铁路车轴的疲劳强度及剩余寿命评价具有重要意义.     

EGCG通过TGF-β1信号通路抑制PLA-802细胞的机制研究

目的 观察表没食子儿茶素-3-没食子酸酯(EGCG)对人横纹肌肉瘤细胞株PLA-802的抑制作用,以及对细胞内TGF-β1/Smad4的表达的影响,探索EGCG抑制横纹肌肉瘤细胞生长的机制.方法 体外培养人横纹肌肉瘤细胞株PLA-802,并用不同浓度的EGCG作用不同的时间.用MTT检测EGCG对PLA-802细胞生长情况的影响,用流式细胞术检测细胞周期的变化情况,RT-PCR和Westernblot分别检测细胞内TGF-β1和Smad4的mRNA和蛋白水平的表达.结果 MTT结果显示EGCG显著降低了PLA-802细胞的存活率((P＜0.05).流式细胞结果表明EGCG明显降低了S期而增加了G1期(P＜0.05).而TGF-β1和其下游因子Smad4的mRNA和蛋白水平的表达也明显受到EGCG的抑制,且这种抑制作用呈浓度-时间依赖性(P＜0.05).结论 EGCG发挥其抑制PLA-802细胞的作用可通过抑制TGF-β1信号通路,这或许将为临床治疗横纹肌肉瘤提供新的思路.     

白菜病害生防原生质体融合菌株的筛选

为了获得同时具有白菜软腐病和黑斑病生防功能的原生质体融合菌株,对白菜软腐病生防菌株芽孢杆菌KR和白菜黑斑病生防菌株假单胞菌B13的融合子进行了筛选.结果表明:2.0单位/mL的硫酸妥布霉素和5×10-3 g/mL的氯霉素可分别抑制假单胞茵B13和芽孢杆菌KR,培养基中添加这两种抗生素可选出融合子.对得到的182个融合子进行传代,连续传10代后,得到一株能稳定遗传的融合子KB1.     

刚地弓形虫培养上清抑制THP-1细胞增殖及诱导凋亡的研究

摘要：目的提取弓形虫体外细胞共培养上清,并研究上清对人急性单核细胞白血病细胞THP-1增殖及凋亡的影响。方法收集对数生长期的THP-1细胞以5X10^7／ml细胞浓度接种于不同培养瓶中,对照组加入含10％胎牛血清的RPMll640,实验组加入相同体积不同数量（2×10^7／ml、4X10^7／ml、8×10^7／m1）弓形虫速殖子培养上清,采用四甲基氮噻唑蓝（MTY）法检测吸光度（A490值）并计算THP-1细胞增殖抑制率;倒置显微镜下观察细胞形态变化;Annexin-V-FITC／PI染色细胞后上流式细胞仪检测各个时间点细胞凋亡率变化,以Western印迹方法分析凋亡相关蛋白Bax、Bcl-2的表达或活性。结果MTY法检测结果弓形虫培养上清呈时间剂量依赖性抑制THP-1细胞株增殖,倒置显微镜下观察处理组细胞有发泡现象和凋亡小体出现。流式细胞仪检测弓形虫感染后的THP-1细胞凋亡率较对照组有升高趋势（P〈0．05）,呈量效依赖性,Westernblot检测刚地弓形虫培养上清作用于THP-l细胞48h后实验组的Bax、Bcl-2蛋白表达较对照组的比值分别有明显的升高与降低（P〈0．05）。结论刚地弓形虫速殖子培养上清对体外培养THP-l细胞增殖有明显的抑制作用,并可诱导THP-1细胞凋亡。     

褐藻酸钠不同脱色方法的比较研究

以海带为原料提取褐藻酸钠,并对褐藻酸钠的脱色方法进行研究.分别采用活性碳、次氯酸钠、过氧化氢和大孔树脂NAR-9、ADS-22、ADS-8对褐藻酸钠进行脱色.通过对多糖脱色率比较发现,四种方法对褐藻酸钠均有脱色效果.次氯酸钠脱色率较高但多糖保留率较低,树脂脱色法明显优于次氯酸钠、过氧化氢和活性发.     

茶多酚对裸鼠喉鳞癌的抑制作用及机制研究

目的观察萘多酚对裸鼠喉鳞癌的抑制作用，并初步探讨其机制。方法将接种Hep22细胞后的裸鼠随机分为4组，其中对照组8只，每天给予生理盐水1ml灌胃；茶多酚高、中、低剂量组各8只，每天分莉给予茶多酚80、160、320mg／kg。20d后处死。分离肿瘤测量其体积，real—time RT—PCR法检测肿癌组织中Src、HK Ⅱ mRNA的表这。己糖激酶活性测定试剂盒测定肿瘤组织中己糖激酶活性。结果茶多酚能硬显降低肿瘤体积，降低肿瘤组织中Src、HK Ⅱ mRNA的表达和己糖激酶活性。结论茶多酚能抑制裸鼠喉鳞癌的生长，其机制可能与抑制肿瘤糖酵解有关。     

Inhibitory effect of flavonoids on DNA-dependent DNA and RNA polymerases

Flavonoids, (-)-epigallocatechin (1), myricetin (2) and quercetin (3), were investigated for inhibitory effects on E. coli DNA polymerase I and T7 bacteriophage RNA polymerase. In both DNA and RNA synthesis, 1 and 3 inhibited enzyme reactions by non-competitive and mixed type inhibition respectively, with regard to template DNAs. Myricetin (2) inhibited DNA and RNA polymerase reactions by mixed type and competitive type inhibition, respectively, with template DNAs. It was suggested that 2 interacts with covalently closed circular DNA.     

Glycoprotein IIb/IIIa blockade inhibits platelet aminophospholipid exposure by potentiating translocase and attenuating scramblase activity

The present study investigated the mechanisms underlying the inhibition of platelet phosphatidylserine (PS) exposure by GPIIb/IIIa blockade. Platelet PS exposure induced by thrombin stimulation was cell-cell contact dependent. GPIIb/IIIa blockade by c7E3 or SR121566 inhibited thrombin-induced platelet PS exposure. Thrombin stimulation induced mild, while A23187 induced extensive platelet-derived microparticle (PDMP) generation. Thrombin-induced PDMP generation was not inhibited by GPIIb/IIIa blockade. Aminophospholipid translocase activity was reduced upon platelet activation by thrombin. The reduction of non-PS-exposing platelets was attenuated by GPIIb/IIIa blockade, while little translocase activity was seen in PS-exposing platelets. Thrombin increased scramblase activity slightly in non-PS-exposing platelets, which was inhibited by GPIIb/IIIa blockade, and markedly enhanced scramblase activity in PS-exposing platelets. Activation of platelet calpain and caspase-3 or cytosolic calcium mobilization were not altered by GPIIb/IIIa inhibition. Thus, GPIIb/IIIa blockade inhibits platelet PS exposure by enhancing translocase activity and attenuating scramblase activity, but does not inhibit PDMP generation. Received 13 December 2006; received after revision 5 February 2007; accepted 9 March 2007     

Effects of mistletoe (Viscum album L.) extracts on cultured tumor cells

Bacterially fermented mistletoe preparations (BFMP) were tested on rat hepatoma tissue culture (HTC) cells and human leukemia Molt 4 cells. A dose-dependent inhibition of the growth rate of the cells was observed. For both cell lines, cytostatic concentrations, expressed in weight of fresh plant, were 0.5 mg/ml culture medium for oak BFMP and 1 mg/ml for apple tree BFMP. However, the action of the two preparations was markedly different on each cell line. Non-viable HTC cells were not stained by trypan blue while non-viable Molt 4 cells were fully colored by this reagent. A lysis of cellular membranes of HTC cells was observed by electron microscopy. Furthermore, oak BFMP inhibited the growth of virus transformed 3T3-SV40 cells more than that of non-transformed 3T3 cells. In contrast to BFMP, non-fermented extracts and a purified mistletoe lectin showed a greater inhibition of the growth of Molt 4 cells than of HTC cells. Samples withdrawn at different times during fermentation gradually lost their inhibitory effect on the growth of Molt 4 cells while their action on HTC cells increased up to the 4th day of fermentation. These results are discussed in relation to the cytotoxic substances of mistletoe already characterized.     

Nucleotide sequence of the gene for the mitochondrial 15S ribosomal RNA of yeast

We have determined the nucleotide sequence of a DNA segment carrying the entire 15S ribosomal RNA gene of yeast mitochondrial genome. Many stretches of sequence are present which are homologous to the E. coli 16S ribosomal RNA gene. The gene sequence can be folded into a secondary structure according to the [1] model on bacterial ribosomal RNAs. The structure reveals a striking similarity between the two RNAs despite the large difference in their base compositions. In the middle of the gene, we found a guanine-cytosine rich sequence that is also present in several other regions of the mitochondrial genome.     

BRD7 regulates the insulin-signaling pathway by increasing phosphorylation of GSK3β

Reduced hepatic expression levels of bromodomain-containing protein 7 (BRD7) have been suggested to play a role in the development of glucose intolerance in obesity. However, the molecular mechanism by which BRD7 regulates glucose metabolism has remained unclear. Here, we show that BRD7 increases phosphorylation of glycogen synthase kinase 3β (GSK3β) in response to activation of the insulin receptor-signaling pathway shortly after insulin stimulation and the nutrient-sensing pathway after feeding. BRD7 mediates phosphorylation of GSK3β at the Serine 9 residue and this effect on GSK3β occurs even in the absence of AKT activity. Using both in vitro and in vivo models, we further demonstrate that BRD7 mediates phosphorylation of ribosomal protein S6 kinase (S6K) and leads to increased phosphorylation of the eukaryotic translation initiation factor 4E-binding protein 1 (4E-BP1) and, therefore, relieves its inhibition of the eukaryotic translation initiation factor 4E (eIF4E). However, the increase in phosphorylation of 4E-BP1 with BRD7 overexpression is blunted in the absence of AKT activity. In addition, using liver-specific BRD7 knockout (LBKO) mice, we show that BRD7 is required for mTORC1 activity on its downstream molecules. These findings show a novel basis for understanding the molecular dynamics of glucose metabolism and suggest the unique function of BRD7 in the regulation of glucose homeostasis.     

Action of iodine on the tomato pectinesterase

Summary The organophosphates and carbamates did not inhibit the isolated tomato pectinesterase. Therefore this enzyme cannot be defined as serine-type esterase. The enzyme is inhibited by iodine and the inhibition (irreversible and non-competitive) is dependent on the degree of enzyme purification.     

Participation of the second extracellular loop of claudin-5 in paracellular tightening against ions, small and large molecules

Tight junctions control paracellular permeability. Here, we analyzed the impact of residues in the second extracellular loop (ECL2) of mouse claudin-5 on paracellular permeability. Stable expression of claudin-5_{wild type} in MDCK-II cells—but not that of mutants R145A, Y148A, Y158A or E159Q—increased transepithelial electrical resistance and decreased fluorescein permeation. Expression of claudin-5_Y148A, _Y158A or _E159Q enhanced permeability of FITC-dextran_10 kDa, which was unchanged in cells expressing claudin-5_{wild type} or claudin-5_R145A. In contrast, targeting to tight junctions, strand morphology and tight junction assembly were unchanged. It is concluded that R145 is unessential for trans-interaction of claudin-5, but necessary for tightening against small solutes and ions. The highly conserved residues Y148, Y158 and E159 in ECL2 of claudin-5 contribute to homo- and/or heterophilic trans-interaction between classic claudins and thereby tighten the paracellular space against ions, small and large molecules. These results provide novel insights into the molecular function of tight junctions.     

Degradation of ribosomal RNA from Escherichia coli by ribonuclease U2 and in vitro transcription of "RNA-fragments" into complementary DNA]

Under well defined conditions ribosomal RNAs purified from Escherichia coli can be degraded by ribonuclease U2 giving rise to RNA fragments of 60--70 nucleotides. In vitro, these fragments are efficiently transcribed into a complementary DNA by DNA polymerase RNA dependent, partially purified from extracts of E. coli. In vivo, "RNA-fragments-U2" inhibit the development of plant tumors.     

Influence of the carbohydrate moiety on the growth inhibitory activity and adhesiveness of 3T3 cell plasma membranes

Summary Treatment of 3T3 cell plasma membranes with glycosidase enzymes decreased their ability to inhibit cell growth and also decreased their binding to 3T3 cells. This suggests that carbohydrate is required for complete function of inhibitory activity and that inhibition is associated with membrane adhesion.     

Influence of the carbohydrate moiety on the growth inhibitory activity and adhesiveness of 3T3 cell plasma membranes

Treatment of 3T3 cell plasma membranes with glycosidase enzymes decreased their ability to inhibit cell growth and also decreased their binding to 3T3 cells. This suggests that carbohydrate is required for complete function of inhibitory activity and that inhibition is associated with membrane adhesion.