小鼠耐力训练后再力竭运动对体内某些生化指标的影响

目的 :通过分析耐力训练后再力竭运动小鼠部分生化指标的变化来探讨耐力训练提高机体抗疲劳能力可能机制及血液再分配的机制。方法 :建立耐力训练后再力竭运动模型 ,测定血清中超氧化物歧化酶 (SOD)活性、过氧化物酶 (POD)活性、过氧化氢酶 (CAT)活性与丙二醛 (MDA )含量及肝、骨骼肌、心肌与血清中一氧化氮 (NO )含量。结果 :运动后即刻 ,非耐力组血清SOD活性及肝NO含量较安静组显著下降 (P <0 .0 5～ 0 .0 1) ,而血清POD与CAT活性及骨骼肌与血清NO含量则显著升高 (P <0 .0 5～ 0 .0 1) ,其余指标无显著变化 (P >0 .0 5)。耐力组CAT活性显著高于非耐力组 (P <0 .0 5) ,血清NO含量显著低于非耐力组 (P <0 .0 1) ,其余指标两组间无显著差异 (P >0 .0 5)。 2 4h恢复后 ,非耐力组血清CAT活性与MDA含量及肝NO较运动后即刻显著升高 (P <0 .0 5～ 0 .0 1) ,血清与骨骼肌NO含量显著下降 (P <0 .0 5) ,而其余指标则无显著变化 (P >0 .0 5)。耐力组血清SOD活性及肝、血清与心肌NO含量较运动后即刻显著升高 (P <0 .0 5) ,而血清CAT活性显著下降 (P <0 .0 5) ,其余指标无显著变化(P >0 .0 5)。耐力组血清CAT活性与MDA含量显著低于非耐力组 (P <0 .0 5) ,而心肌NO含量显著高于非耐力组 (P <0 .0 5) ,其余指标两组间     

硅对污灌水稻幼苗几种生理特性的影响

水稻幼苗经奎河污水胁迫后 ,根系活力、叶绿素含量、叶绿素 a/b比值下降 ;叶片 POD活性、SOD活性、相对电导率升高。在污水中加入 Si( 1 .5 mmol/L) ,可使前三项生理指标明显提高 ( P<0 .0 1 ) ,而后三项生理指标明显下降 ( P<0 .0 1 )。说明 Si在一定程度上可以减轻奎河污水对水稻幼苗的毒害作用。     

脓毒血症早晚期肝细胞核被膜NTPase活性及poly(A)+mRNA核浆转运的变化

目的和方法 :在盲肠结扎穿孔脓毒血症大鼠模型上 ,研究早晚期脓毒血症肝细胞核被膜核苷三磷酸酶 (NT Pase)活性及 poly(A) mRNA核浆转运的变化。结果 :脓毒血症早期NTPase活性增加而晚期活性降低 ,即早期Vmax与对照组比增加 6 2 % (ATP ,P <0 .0 5 )和 18% (GTP ,P <0 .0 5 ) ;晚期Vmax下降 2 6 % (ATP ,P <0 .0 5 )和5 6 % (GTP ,P <0 .0 5 )。脓毒血症仅晚期NTPase底物亲和力降低 ,即晚期Km与对照组比分别升高 2 6 % (ATP ,P<0 .0 5 )和 37% (GTP ,P <0 .0 5 )。脓毒血症早期 poly(A) mRNA核浆转运速率增加而晚期降低 ,即早期组 10min转运速率较对照组增加 2 7% (ATP ,P <0 .0 5 )和 30 % (GTP ,P <0 .0 5 ) ;晚期组降低 2 1% (P <0 .0 1)和 35 % (P <0 .0 1)。结论 :脓毒血症肝细胞核膜NTPase活性呈早期升高、晚期下降的双相变化与肝细胞核被膜 poly(A) mR NA核浆转运速率的早、晚期变化相平行 ,同时与脓毒血症血糖浓度的早、晚期变化相关。本实验在一定程度上解释了脓毒血症早晚期代谢双相变化发生的分子机理     

竹叶青蛇伤血液变化特点及处理

目的　评价竹叶青蛇伤后血液变化特点 ,总结处理方法。　方法　对被诊断为竹叶青蛇伤的 42例患者作前瞻性血液系统功能检查。　结果　 8例血小板低于正常 ,最低 1 0× 1 0 9/L,纤维蛋白原 (1 .66± 0 .5 9) g/L(P <0 .0 1 ) ,其中 3例降低至测不出 ,PT活性 (61 .66± 8.61 ) % ,凝血酶原时间 (3 0 .2 3±9.3 8) s(P <0 .0 5 ) ,PT比率 (2 .5 3± 0 .96) (P<0 .0 5 ) ,部分凝血活酶时间 (5 1 .92± 2 1 .63 ) s(P<0 .0 5 ) ,PTT比率 (1 .2 5± 0 .2 8) (P <0 .0 5 ) ,凝血酶时间 (4 0 .1 7± 1 2 .42 ) s(P <0 .0 5 ) ;8例 3 P试验阳性 ,7例D-二聚体阳性。　结论　竹叶青蛇伤血液变化普遍呈纤维蛋白原下降 ,凝血功能下降。血小板减少程度可帮助判断病情危重。     

硒元素对平菇菌丝体GSH-Px、SOD及MDA的影响

于培养基中加入一定量的亚硒酸钠溶液 ,分别测定了 2个品种平菇菌丝体内GSH Px、SOD活性及MDA含量。结果表明 :3 0、60mg/L组菌丝体内GSH Px、SOD活性极显著升高 (P <0 .0 1 )而MDA含量明显降低 (P <0 .0 5 ) ,随着硒水平的升高 ,GSH Px、SOD活性呈下降趋势而MDA含量则显著升高 (P <0 .0 5 )。因此 ,在培养富硒平菇菌丝体时应适当考虑培养基的硒浓度。     

凝血因子FⅧA链基因第1内含子内顺式调节元件的鉴定

探讨FⅧA基因表达的分子机制.凝胶阻滞实验(EMSA)结果证明,FⅧA基因第1内含子5′区的前12碱基与转录因子结合并由此调控基因表达.FⅧA基因第1内含子第12位碱基由C突变为A(内含子1( 12)C→A)导致与转录因子结合能力降低.构建不同的荧光素酶表达质粒Luc1、Luc2、Luc3、Luc4、Luc5、Luc6,并转染U937细胞和HepG2细胞.结果显示,如果Luc5(具有最高表达活性)的内含子1( 12)由C突变为A,启动子活性显著降低.与Luc5相比,突变后的Luc5的活性分别下降了52·9%(U937,P<0·01)和47·6%(HepG2,P<0·01).将Luc5与PN3 Sp1共同转染U937细胞和HepG2细胞后,Luc5的荧光素酶活性分别提高了42·4%(U937,与Luc5单独转染比较P<0·01)和54·9%(HepG2,与Luc5单独转染比较P<0·01),而突变后的Luc5(Mut)与PN3 Sp1共同转染则没有明显的改变.表明转录因子Sp1在FXA基因表达的重要性.这些结果也表明,内含子在FⅧA基因表达过程中起重要作用,为遗传性凝血因子Ⅷ发病机理的研究提供了新的证据.     

抗Caspase-3核酶M1RNA的制备与体内外活性鉴定

为评价抗caspase 3核酶在阻抑细胞凋亡发生中的潜在价值 ,以RNaseP催化亚基M1RNA为模板 ,设计合成 3个特异性针对人caspase 3的核酶pM1 GS716、pM1 GS337和pM1 GS2 35 ,并对它们的体内外切割活性进行探讨 .3 2 P标记的caspase 3基因片段体外转录物作为靶RNA ,体外切割实验表明 ,pM1 GS716和pM1 GS337均有切割活性 ,其中pM1 GS716的切割效率可达到 93% .3个核酶转染HeLa细胞 ,评价其在体内的切割活性 .在TNF α作用下 ,转染pM1 GS716的HeLa细胞内caspase 3mRNA下降了 75 % ,蛋白含量下降了 6 9% ,caspase 3蛋白酶活性下降了 5 2 % .Hoechst 332 5 8染色表明 ,细胞凋亡率较对照明显下降 (分别为 2 1 6± 0 7%和 4 9 4± 0 2 % ,P <0 0 1) .提示体外制备的pM1 GS716具有良好的特异催化切割活性 ,有望通过切割caspase 3而抑制细胞凋亡 .     

不同氮源对苦草(Vallisneria natans)生长及生理指标的影响

通过实验室静态模拟,研究了在富营养条件下(4.0 mg.L-1TN,0.2 mg.L-1TP)不同比例铵态氮和硝态氮(6∶0、5∶1、3∶3、1∶5和0∶6)对苦草〔Vallisneria natans(Lour.)Hara〕的生长与生理指标的影响。结果表明,随着铵态氮和硝态氮比例的下降,苦草相对生长率和蛋白质含量先升高后下降,超氧化物歧化酶(SOD)和谷氨酰胺合成酶(GS)活性逐渐下降,硝酸还原酶(NR)活性逐渐升高。在4.0 mg.L-1TN和0.2 mg.L-1TP条件下,若不考虑磷的作用,高浓度铵态氮对苦草的生理功能有影响,对其生长有明显的抑制作用;而当铵态氮浓度小于0.67 mg.L-1时却可以促进苦草的生长。     

服用CH-1双歧杆菌制剂对I型糖尿病患者、Ⅱ型糖尿病患者以及透析患者肠内细菌群以及生化指标影响的研究

随机抽取 型、 型糖尿病以及透析患者各 10例进行 CH- 1双歧杆菌制剂服用试验。并对前述患者服用 CH - 1双歧杆菌制剂的肠内细菌群和肠内腐败物质以及生化指标进行检测分析。 型糖尿病患者服用期间 ,双歧杆菌数占肠内总菌数的百分率由 (0 .84± 0 .75 )‰升高到服用 14 d后的 (7.2 5± 7.4 9)‰ ,差异有显著性。腐败菌数亦显著下降 ,卵磷脂酶阳性腐败梭菌由 8.14± 0 .37下降到服用 14 d后的 6 .0 9±0 .4 9(差异有十分显著性 )。 型糖尿病患者服用期间 ,双歧杆菌数占肠内总菌数的百分率由 (2 .4 0±3.18)‰升高到服用 14 d后的 (5 .5 6± 7.4 8)‰ ,差异有显著性。腐败菌数亦显著下降 ,卵膦脂酶阳性腐败梭菌由 7.99± 1.15下降到服用 14 d后的 5 .93± 0 .17(差异有十分显著性 )。透析患者患者服用期间 ,双歧杆菌数占肠内总菌数的百分率由 (0 .6 78± 0 .92 )‰升高到服用 14 d后的 (7.0 8± 6 .95 )‰ ,差异有显著性。腐败菌数亦显著下降 ,卵膦脂酶阳性腐败梭菌由 8.33± 0 .5 0下降到服用 14 d后的 5 .96± 0 .4 4 (差异有十分显著性 )。服用 CH- 1双歧杆菌制剂后粪便中腐败物质含量的变化 , 型糖尿病患者服用 CH- 1双歧杆菌制剂后 ,氨由 82 3± 76 .2 (μg/ g)下降到服用 14 d后的 4 71± 73.0 ,     

MKL1靶向CAAP1促进胃癌细胞的自噬并抑制凋亡

[目的]探讨MKL1和CAAP1对胃癌细胞凋亡的影响及其分子机制。[方法]蛋白质印迹和实时荧光定量PCR检测MKL1、CAAP1过表达效果;蛋白质印迹、流式细胞术、自噬检测试剂盒检测细胞的自噬和凋亡情况;荧光素酶报告基因活性测定、染色质免疫共沉淀验证MKL1和CAAP1基因启动子的结合。[结果]在MGC80-3细胞中过量表达MKL1后,凋亡抑制蛋白CAAP1的mRNA和蛋白水平与对照组相比均上调(P 0. 05),自噬标记基因LC3BII表达升高(P 0. 05),促凋亡蛋白Caspase3表达降低(P 0. 05)。敲降MKL1,Caspase3蛋白表达升高(P 0. 05)。敲降CAAP1,Caspase3蛋白表达升高(P 0. 05)。过表达CAAP1拮抗敲降MKL1,LC3BII蛋白表达下降,Caspase3蛋白表达升高(P 0. 05)。荧光素酶活性分析以及染色质免疫共沉淀分析结果表明MKL1促进CAAP1基因的转录活性。[结论]MKL1可通过上调CAAP1的表达进而促进胃癌细胞的自噬并抑制细胞凋亡。     

LINE-1编码蛋白L1-ORF1的原核表达纯化和多克隆抗体制备

目的: 制备具有肿瘤组织特异性表达的L1-ORF1蛋白多克隆抗体并进行初步应用研究。方法:采取基因工程表达方法制备L1-ORF1蛋白,免疫家兔制备多克隆抗体,间接ELISA检测抗体效价,Western blot和细胞免疫荧光方法检测抗体特异性,免疫检测验证其识别肿瘤细胞内L1-ORF1蛋白的特异性。结果:制备的抗L1-ORF1蛋白多克隆抗体具有很高的敏感性与特异性,免疫学检测表明该抗体不仅能检测出正常细胞中瞬时表达的L1-ORF1蛋白,而且可检测出肿瘤细胞中天然表达的L1-ORF1蛋白。结论:制备的多克隆抗体具有较高的敏感性与特异性,为以后该抗体的进一步应用奠定了基础。     

Role of the ASK1-SEK1-JNK1-HIPK1 signal in Daxx trafficking and ASK1 oligomerization

Overexpression of JNK binding domain inhibited glucose deprivation-induced JNK1 activation, relocalization of Daxx from the nucleus to the cytoplasm, and apoptosis signal-regulating kinase 1 (ASK1) oligomerization in human prostate adenocarcinoma DU-145 cells. However, SB203580, a p38 inhibitor, did not prevent relocalization of Daxx and oligomerization of ASK1 during glucose deprivation. Studies from in vivo labeling and immune complex kinase assay demonstrated that phosphorylation of Daxx occurred during glucose deprivation, and its phosphorylation was mediated through the ASK1-SEK1-JNK1-HIPK1 signal transduction pathway. Data from immunofluorescence staining and protein interaction assay suggest that phosphorylated Daxx may be translocated to the cytoplasm, bind to ASK1, and subsequently lead to ASK1 oligomerization. Mutation of Daxx Ser667 to Ala results in suppression of Daxx relocalization during glucose deprivation, suggesting that Ser667 residue plays an important role in the relocalization of Daxx. Unlike wild-type Daxx, a Daxx deletion mutant (amino acids 501-625) mainly localized to the cytoplasm, where it associated with ASK1, activated JNK1, and induced ASK1 oligomerization without glucose deprivation. Taken together, these results show that glucose deprivation activates the ASK1-SEK1-JNK1-HIPK1 pathway, and the activated HIPK1 is probably involved in the relocalization of Daxx from the nucleus to the cytoplasm. The relocalized Daxx may play an important role in glucose deprivation-induced ASK1 oligomerization.     

Antagonism between Cry1Ac1 and Cyt1A1 toxins of bacillus thuringiensis

Most strains of the insecticidal bacterium Bacillus thuringiensis have a combination of different protoxins in their parasporal crystals. Some of the combinations clearly interact synergistically, like the toxins present in B. thuringiensis subsp. israelensis. In this paper we describe a novel joint activity of toxins from different strains of B. thuringiensis. In vitro bioassays in which we used pure, trypsin-activated Cry1Ac1 proteins from B. thuringiensis subsp. kurstaki, Cyt1A1 from B. thuringiensis subsp. israelensis, and Trichoplusia ni BTI-Tn5B1-4 cells revealed contrasting susceptibility characteristics. The 50% lethal concentrations (LC50s) were estimated to be 4,967 of Cry1Ac1 per ml of medium and 11.69 ng of Cyt1A1 per ml of medium. When mixtures of these toxins in different proportions were assayed, eight different LC50s were obtained. All of these LC50s were significantly higher than the expected LC50s of the mixtures. In addition, a series of bioassays were performed with late first-instar larvae of the cabbage looper and pure Cry1Ac1 and Cyt1A1 crystals, as well as two different combinations of the two toxins. The estimated mean LC50 of Cry1Ac1 was 2.46 ng/cm2 of diet, while Cyt1A1 crystals exhibited no toxicity, even at very high concentrations. The estimated mean LC50s of Cry1Ac1 crystals were 15.69 and 19.05 ng per cm2 of diet when these crystals were mixed with 100 and 1,000 ng of Cyt1A1 crystals per cm2 of diet, respectively. These results indicate that there is clear antagonism between the two toxins both in vitro and in vivo. Other joint-action analyses corroborated these results. Although this is the second report of antagonism between B. thuringiensis toxins, our evidence is the first evidence of antagonism between toxins from different subspecies of B. thuringiensis (B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. israelensis) detected both in vivo and in vitro. Some possible explanations for this relationship are discussed.     

Isolation and Characterization of Ubiquitin-activating Enzyme E1-domain Containing 1, UBE1DC1

Ubiquitin and other ubiquitin-like proteins play important roles in post-translational modification. They are phylogenetically well-conserved in eukaryotes. Activated by other proteins, ubiquitin and ubiquitin-like proteins can covalently modify target proteins. The enzymes responsible for the activation of this modification have been known to include UBA1, SAE2, UBA3, SAE1 and ULA1. Here we report a new ubiquitin activating enzyme like cDNA, named ubiquitin activating enzyme E1-domain containing 1 (UBE1DC1), whose cDNA is 2654 base pairs in length and contains an open reading frame encoding 404 amino acids. The UBE1DC1 gene consists of 12 exons and is located at human chromosome 3q22. The result of RT-PCR showed that UBE1DC1 is expressed in most of human tissues. These two authors contributed equally to this paper. The nucleotide sequence reported in this paper has been submitted to GenBank under accession number AY253672.     

Regulation of PCTAIRE1 protein stability by AKT1, LKB1 and BRCA1

PCTAIRE1, also known as CDK16, is a cyclin-dependent kinase that is regulated by cyclin Y. It is a member of the serine-threonine family of kinases and its functions have primarily been implicated in cellular processes like vesicular transport, neuronal growth and development, myogenesis, spermatogenesis and cell proliferation. However, as extensive studies on PCTAIRE1 have not yet been conducted, the signaling pathways for this kinase involved in governing many cellular processes are yet to be elucidated in detail. Here, we report the association of PCTAIRE1 with important cellular proteins involved in major cell signaling pathways, especially cell proliferation. In particular, here we show that PCTAIRE1 interacts with AKT1, a key player of the PI3K signaling pathway that is responsible for promoting cell survival and proliferation. Our studies show that PCTAIRE1 is a substrate of AKT1 that gets stabilized by it. Further, we show that PCTAIRE1 also interacts with and is degraded by LKB1, a kinase that is known to suppress cellular proliferation and also regulate cellular energy metabolism. Moreover, our results show that PCTAIRE1 is also degraded by BRCA1, a well-known tumor suppressor. Together, our studies highlight the regulation of PCTAIRE1 by key players of the major cell signaling pathways involved in regulating cell proliferation, and therefore, provide crucial links that could be explored further to elucidate the mechanistic role of PCTAIRE1 in cell proliferation and tumorigenesis.     

Antagonism between Cry1Ac1 and Cyt1A1 Toxins of Bacillus thuringiensis

Most strains of the insecticidal bacterium Bacillus thuringiensis have a combination of different protoxins in their parasporal crystals. Some of the combinations clearly interact synergistically, like the toxins present in B. thuringiensis subsp. israelensis. In this paper we describe a novel joint activity of toxins from different strains of B. thuringiensis. In vitro bioassays in which we used pure, trypsin-activated Cry1Ac1 proteins from B. thuringiensis subsp. kurstaki, Cyt1A1 from B. thuringiensis subsp. israelensis, and Trichoplusia ni BTI-Tn5B1-4 cells revealed contrasting susceptibility characteristics. The 50% lethal concentrations (LC₅₀s) were estimated to be 4,967 of Cry1Ac1 per ml of medium and 11.69 ng of Cyt1A1 per ml of medium. When mixtures of these toxins in different proportions were assayed, eight different LC₅₀s were obtained. All of these LC₅₀s were significantly higher than the expected LC₅₀s of the mixtures. In addition, a series of bioassays were performed with late first-instar larvae of the cabbage looper and pure Cry1Ac1 and Cyt1A1 crystals, as well as two different combinations of the two toxins. The estimated mean LC₅₀ of Cry1Ac1 was 2.46 ng/cm² of diet, while Cyt1A1 crystals exhibited no toxicity, even at very high concentrations. The estimated mean LC₅₀s of Cry1Ac1 crystals were 15.69 and 19.05 ng per cm² of diet when these crystals were mixed with 100 and 1,000 ng of Cyt1A1 crystals per cm² of diet, respectively. These results indicate that there is clear antagonism between the two toxins both in vitro and in vivo. Other joint-action analyses corroborated these results. Although this is the second report of antagonism between B. thuringiensis toxins, our evidence is the first evidence of antagonism between toxins from different subspecies of B. thuringiensis (B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. israelensis) detected both in vivo and in vitro. Some possible explanations for this relationship are discussed.     

Synthesis of 1-benzyl-1H-benzimidazoles as galectin-1 mediated anticancer agents

In our pursuit to develop novel non-carbohydrate small molecule Galectin-1 Inhibitors, we have designed a series of 1-benzyl-1H-benzimidazole derivatives and demonstrated their anticancer activity. The compound 6g, 4-(1-benzyl-5-chloro-1H-benzo[d]imidazol-2-yl)-N-(4-hydroxyphenyl) benzamide was found to be most potent with an IC₅₀ of 7.01 ± 0.20 µM and arresting MCF-7 cell growth at G2/M phase and S phase. Induction of apoptosis was confirmed by morphological changes like cell shrinkage, blebbing and cell wall deformation, dose dependent increase in the mitochondrial membrane potential (ΔΨm) and ROS levels. Further, dose dependent decrease in Gal-1 protein levels proves Gal-1 mediated apoptosis by 6g. Molecular docking studies were performed to understand the Gal-1 interaction with compound 6g. In addition, RP-HPLC studies showed 85.44% of 6g binding to Gal-1. Binding affinity studies by fluorescence spectroscopy and Surface Plasmon Resonance (SPR) showed that 6g binds to Gal-1 with binding constant (K_a) of 1.2 × 10⁴ M⁻¹ and equilibrium constant K_D value of 5.76 × 10⁻⁴ M respectively.