• 1 Bioneedles as vaccine carriers. 复制标题 收藏 收藏

    【Bioneedles作为疫苗载体。】 复制标题 收藏 收藏
    DOI:10.1016/j.vaccine.2008.02.067 复制DOI
    作者列表:Hirschberg HJ,van de Wijdeven GG,Kelder AB,van den Dobbelsteen GP,Kersten GF
    BACKGROUND & AIMS: :Bioneedles are small hollow mini implants fabricated from biodegradable polymers which can be filled with antigen. Bioneedles can be used for vaccination without syringes and needles. Formulations have been prepared containing tetanus toxoid with and without aluminum phosphate. Stability and immunogenicity of Bioneedles were compared with liquid formulations. The antigen, when formulated in Bioneedles, retained fully its antigenicity up to 60 degrees C for 1 week whereas the antigen, in its liquid form, lost all activity at 60 degrees C after 1 week. After 3 weeks at 60 degrees C, a recovery of 60% was still found in the Bioneedles. Mice injected with Bioneedles with adjuvanted tetanus toxoid showed a comparable functional antibody response to the group receiving conventional liquid injections. This response was achieved with a four times lower antigen concentration when using the Bioneedles compared to the regular injections. We conclude that Bioneedles are good alternatives for injections using needles and syringes.
    背景与目标: :Bioneedles是由可生物降解的聚合物制成的小型空心微型植入物,其中可充满抗原。不用注射器和针头也可以使用Bioneedles进行疫苗接种。已经制备了含有破伤风类毒素且具有和不具有磷酸铝的制剂。将Bioneedles的稳定性和免疫原性与液体制剂进行了比较。当在Bioneedles中配制抗原时,在60摄氏度以下的温度下可完全保留其抗原性1周,而液体形式的抗原在60摄氏度下的1周后会失去所有活性。在60摄氏度下3周后,在Bioneedles中仍发现60%的回收率。注射了带助剂破伤风类毒素的双甲醚的小鼠显示出与接受常规液体注射的组相当的功能抗体反应。与常规注射相比,使用Bioneedles时,抗原浓度降低了四倍,从而达到了这种反应。我们得出结论,Bioneedles是使用针头和注射器进行注射的好选择。
  • 【成人T细胞白血病和健康HTLV-1携带者的细胞骨架系统血清抗体。】 复制标题 收藏 收藏
    DOI:10.3109/10428199109068089 复制DOI
    作者列表:Yasuda M,Nobunaga M
    BACKGROUND & AIMS: :Serum antibodies to cytoskeletal systems were detected, using indirect immunofluorescence in patients with adult T-cell leukemia (ATL), healthy carriers of human T cell lymphotropic virus 1 (HTLV-1), patients with infectious mononucleosis and healthy adults. Healthy carriers of HTLV-1 had IgG antibodies to cytoskeletal systems as evidenced by an increased incidence of IgG antibodies to actin and vimentin. Decreased IgG antibody levels to Epstein-Barr virus nucleic acid (EBNA) were also evident. In patients with ATL, the titers of IgM antibodies to vimentin and cytokeratin showed a positive correlation with decreased serum levels of IgM, despite the fact that serum concentrations of IgM were significantly decreased in patients with ATL. The IgM antibody titer divided by the IgM concentration (the antibody ratio) was nigher than that of healthy carriers and healthy adults, suggesting that the IgM antibody response to cytoskeletal systems was preferentially preserved in these cases. There was also a suggestion of the presence of polyclonal as well as specific antibody responses to cytoskeletal systems in patients with infectious mononucleosis. As a result of these findings we suggest that there is some difference in the mechanisms responsible for the production of autoantibodies to cytoskeletal systems following HTLV-1 infection and Epstein-Barr virus infection.
    背景与目标: :在成人T细胞白血病(ATL),健康的人T细胞淋巴病毒1(HTLV-1)携带者,传染性单核细胞增多症患者和健康成年人中,使用间接免疫荧光检测了针对细胞骨架系统的血清抗体。健康的HTLV-1携带者具有针对细胞骨架系统的IgG抗体,这可以通过针对肌动蛋白和波形蛋白的IgG抗体发生率的增加来证明。对爱泼斯坦-巴尔病毒核酸(EBNA)的IgG抗体水平降低也很明显。在ATL患者中,针对波形蛋白和细胞角蛋白的IgM抗体滴度与血清IgM水平降低呈正相关,尽管ATL患者的IgM血清浓度显着降低。 IgM抗体滴度除以IgM浓度(抗体比率)比健康携带者和健康成年人要高,这表明在这些情况下,优先保留了对细胞骨架系统的IgM抗体反应。还提示感染性单核细胞增多症患者存在多克隆以及对细胞骨架系统的特异性抗体反应。这些发现的结果表明,在HTLV-1感染和爱泼斯坦-巴尔病毒感染后,产生针对细胞骨架系统的自身抗体的机制存在一定差异。
  • 【分支的基于聚乙烯亚胺的PKCα反应基因载体。】 复制标题 收藏 收藏
    DOI:10.1080/09205063.2013.807459 复制DOI
    作者列表:Nakamura Y,Kim CW,Tsuchiya A,Kushio S,Nobori T,Li K,Lee EK,Zhao GX,Funamoto D,Niidome T,Mori T,Katayama Y
    BACKGROUND & AIMS: :We examined in vitro performance of the branched polyethylenimine (bPEI)-based gene carriers which respond to cancer-specific activation of protein kinase Cα (PKCα) to express plasmid DNA. The carriers were synthesized straightforward by using amide bond formation between a peptide terminal carboxyl and a primary amine group of bPEI. To examine the effect of the peptide contents in the carrier, we prepared several carriers with various peptide contents. The obtained polymers form polyplexes with tighter condensation of plasmid DNA than our previous gene carriers. After internalization of the polyplexes via endocytosis, the polyplexes effectively escaped from the endosome into cytosol. Then, the polyplexes showed a clear-cut response to PKCα to release plasmid DNA for gene expression. We determined the optimum contents of the peptides in carriers as 5 mol% to achieve the clear-cut response to PKCα.
    背景与目标: :我们研究了分支的基于聚乙烯亚胺(bPEI)的基因载体的体外性能,这些载体对癌症特异性的蛋白激酶Cα(PKCα)活化表达质粒DNA有反应。通过使用肽末端羧基和bPEI的伯胺基团之间的酰胺键形成,直接合成了载体。为了检查载体中肽含量的影响,我们制备了几种具有各种肽含量的载体。与我们以前的基因载体相比,所获得的聚合物形成了质粒DNA紧密结合的多链体。通过胞吞作用使多聚体内在化后,多聚体有效地从内体逃逸到胞质溶胶中。然后,复合物显示出对PKCα的明确反应,释放出质粒DNA用于基因表达。我们确定肽在载体中的最佳含量为5摩尔%,以实现对PKCα的明确反应。
  • 【PGRN单倍体不足会增加额颞叶大叶变性-progranulin突变携带者周围细胞中的Wnt5a信号传导。】 复制标题 收藏 收藏
    DOI:10.1016/j.neurobiolaging.2013.09.021 复制DOI
    作者列表:Alquézar C,Esteras N,de la Encarnación A,Alzualde A,Moreno F,López de Munain A,Martín-Requero A
    BACKGROUND & AIMS: :Loss-of-function progranulin (PGRN) mutations have been identified as the major cause of frontotemporal lobar degeneration with TDP-43 protein inclusions (FTLD-TDP). Previously, we reported cell cycle-related alterations in lymphoblasts from FTLD-TDP patients, carrying the c.709-1G>A null PGRN mutation, suggesting aberrant cell cycle activation in affected neurons. Here we report that PGRN haploinsufficiency activates the extracellular signal-regulated protein kinases 1 and 2 pathway in a Ca(2+), protein kinase C-dependent, and pertussis toxin-sensitive manner. Addition of exogenous PGRN or conditioned medium from control cells normalized the response of PGRN-deficient lymphoblasts to serum activation. Our data indicated that noncanonical Wnt5a signaling might be overactivated by PGRN deficiency. We detected increased cellular and secreted levels of Wnt5a in PGRN-deficient lymphoblasts associated with enhanced phosphorylated calmodulin kinase II. Moreover, treatment of control cells with exogenous Wingless-type 5a (Wnt5a)-activated Ca(2+)/calmodulin kinase II (CaMKII), increased extracellular signal-regulated protein kinases 1 and 2 activity and cell proliferation up to the levels found in c.709-1G>A carrier cells. PGRN knockdown SH-SY5Y neuroblastoma cells also show enhanced Wnt5a content and signaling. Taken together, our results revealed an important role of Wnt signaling in FTLD-TDP pathology and suggest a novel target for therapeutic intervention.
    背景与目标: :功能丧失的前颗粒蛋白(PGRN)突变已被确定为额颞叶大叶变性的主要病因,伴有TDP-43蛋白包涵体(FTLD-TDP)。以前,我们报道了FTLD-TDP患者淋巴母细胞中与细胞周期相关的变化,携带c.709-1G> A无效PGRN突变,表明受影响的神经元中异常的细胞周期激活。在这里我们报告说,PGRN haploinsufficiency激活Ca(2),蛋白激酶C依赖和百日咳毒素敏感方式的细胞外信号调节蛋白激酶1和2途径。从对照细胞中加入外源PGRN或条件培养基使PGRN缺陷的成淋巴细胞对血清活化的反应正常化。我们的数据表明,PGRN缺乏可能会导致非规范的Wnt5a信号过度激活。我们检测到与增强的磷酸化钙调蛋白激酶II相关的PGRN缺陷型淋巴母细胞中Wnt5a的细胞和分泌水平增加。此外,控制细胞与外源性无翼型5a(Wnt5a)激活的Ca(2)/钙调蛋白激酶II(CaMKII)的处理,增加了细胞外信号调节蛋白激酶1和2的活性,并使细胞增殖达到c中所发现的水平.709-1G> A载体细胞。 PGRN组合式SH-SY5Y神经母细胞瘤细胞也显示出增强的Wnt5a含量和信号传导。两者合计,我们的结果揭示了Wnt信号在FTLD-TDP病理学中的重要作用,并提出了治疗干预的新目标。
  • 【脊髓性肌萎缩无症状携带者中生存运动神经元(SMNT)基因缺失的特征。】 复制标题 收藏 收藏
    DOI:10.1093/hmg/5.3.359 复制DOI
    作者列表:Wang CH,Xu J,Carter TA,Ross BM,Dominski MK,Bellcross CA,Penchaszadeh GK,Munsat TL,Gilliam TC
    BACKGROUND & AIMS: Previous reports have established that the telomeric copy of the survival motor neuron (SMNT) gene and the intact copy of the neuronal apoptosis inhibitory protein (NAIP) gene are preferentially deleted in patients with spinal muscular atrophy (SMA). Although deletions or mutations in the SMNT gene are most highly correlated with SMA, it is not clear to what extent NAIP or other genes influence the SMA phenotype, or whether a small fraction of SMA patients actually have functional copies of both SMNT and NAIP. To evaluate further the part of SMNT in the development of SMA, we analyzed 280 asymptomatic SMA family members for the presence or absence of SMNT exons 7 and 8. We report the following observations(i) 4% of the sample harbored a polymorphic variant of SMNT exon 7 that looks like a homozygous deletion; (ii) approximately 1% of the parents are homozygously deleted for both exons 7 and 8; (iii) one asymptomatic parent lacking both copies of SMNT exons 7 and 8 displays a 'subclinical phenotype' characterized by mild neurogenic pathology; (iv) another asymptomatic parent lacking both SMNT exons showed no signs of motor neuron disorder by clinical and neurodiagnostic analyses. The demonstration of polymorphic variants of exon 7 that masquerade as homozygous nulls, and the identification of SMA parents who harbor two disease alleles, serve as a caution to those conducting prenatal tests with these markers.

    背景与目标: 先前的报道已经确定,在脊髓性肌萎缩症(SMA)患者中,优先删除存活运动神经元(SMNT)基因的端粒拷贝和神经元凋亡抑制蛋白(NAIP)基因的完整拷贝。尽管SMNT基因的缺失或突变与SMA高度相关,但尚不清楚NAIP或其他基因在多大程度上影响SMA表型,或一小部分SMA患者实际上是否同时具有SMNT和NAIP的功能性拷贝。为了进一步评估SMNT在SMA发生中的作用,我们分析了280个无症状SMA家族成员是否存在SMNT外显子7和8。看起来像纯合缺失的SMNT外显子7; (ii)对于第7外显子和第8外显子纯合缺失约1%的父母; (iii)缺乏SMNT外显子7和8的两个拷贝的无症状父母表现出一种“亚临床表型”,其特征为轻度神经源性病理; (iv)通过临床和神经诊断分析,另一名既没有SMNT外显子又无症状的父母没有运动神经元疾病的迹象。伪装成纯合无效位点的外显子7多态变异体的演示,以及带有两个疾病等位基因的SMA亲本的鉴定,为那些使用这些标记进行产前检查的人提供了警告。

  • 【一氧化氮合酶抑制剂通过人红细胞中阳离子氨基酸载体的运输。】 复制标题 收藏 收藏
    DOI:10.1016/0006-2952(95)02090-x 复制DOI
    作者列表:Forray MI,Angelo S,Boyd CA,Devés R
    BACKGROUND & AIMS: The interaction of arginine analogues, which are known to inhibit nitric oxide synthase, with two cationic amino acid transporters of human erythrocytes (systems y+ and y+L) was studied. Arginine and relevant analogues [NG-monomethyl-L-arginine (L-NMMA); NG-monomethyl-D-arginine (D-NMMA) and NG-nitro-L-arginine (L-NOARG)] were found to inhibit labeled lysine influx into intact erythrocytes. As expected, the pattern of inhibition reflected the contribution of the two distinct transport systems. All analogues showed a higher affinity for system y+L than for system y+. The half-saturation (inhibition) constants estimated for systems y+ and y+L (+/- SEM) were (microM)L-arginine, 55.7 +/- 5.4 and 2.4 +/- 0.1; L-NMMA, 151 +/- 13 and 7.5 +/- 0.5; D-NMMA, 2660 +/- 404 and 269 +/- 25; L-NOARG, 9414 +/- 169 and 594 +/- 35. The transport properties of the analogues were investigated using an assay based on the trans-stimulation of lysine efflux. The addition of saturating concentrations of unlabeled analogues to the external medium stimulated efflux of labeled lysine through systems y+L and y+, showing that the analogues can enter the cell through these pathways.

    背景与目标: 研究了已知能抑制一氧化氮合酶的精氨酸类似物与人类红细胞的两种阳离子氨基酸转运蛋白(系统y和y L)的相互作用。精氨酸和相关类似物[NG-单甲基-L-精氨酸(L-NMMA);发现NG-单甲基-D-精氨酸(D-NMMA)和NG-硝基-L-精氨酸(L-NOARG)]可抑制标记的赖氨酸流入完整的红细胞。不出所料,抑制模式反映了两种不同运输系统的贡献。所有类似物对系统y L的亲和力均高于对系统y的亲和力。对于系统y和y L(/ -SEM)估计的半饱和(抑制)常数为(microM)L-精氨酸,55.7 / -5.4和2.4 / -0.1; L-NMMA,151 /-13和7.5 /-0.5; D-NMMA,2660 /-404和269 /-25; L-NOARG,9414 /-169和594 /-35。使用基于赖氨酸外排的反式刺激的分析方法研究了类似物的转运特性。将饱和浓度的未标记类似物添加到外部介质中会刺激标记的赖氨酸通过系统y L和y流出,这表明这些类似物可以通过这些途径进入细胞。

  • 【用氨基硅烷修饰的二氧化硅纳米颗粒作为质粒DNA的载体。】 复制标题 收藏 收藏
    DOI:10.1016/s0378-5173(99)00435-4 复制DOI
    作者列表:Kneuer C,Sameti M,Haltner EG,Schiestel T,Schirra H,Schmidt H,Lehr CM
    BACKGROUND & AIMS: :We synthesised silica nanoparticles (SiNP) with covalently linked cationic surface modifications and demonstrated their ability to electrostatically bind, condense and protect plasmid DNA. These particles might be utilised as DNA carriers for gene delivery. All nanoparticles were sized between 10 and 100 nm and displayed surface charge potentials from +7 to +31 mV at pH 7.4. They were produced by modification of commercially available (IPAST) or in-house synthesised silica particles with either N-(2-aminoethyl)-3-aminopropyltrimethoxysilane or N-(6-aminohexyl)-3-aminopropyltrimethoxysilane. All particles formed complexes with pCMVbeta plasmid DNA as evidenced by ratio dependent retardation of DNA in the agarose gel and co-sedimentation of soluble DNA with nanoparticles. High salt and alkaline pH did inhibit complex formation. Absorption onto the particles also decreased the hydrodynamic dimensions of plasmid DNA as shown by photon correlation spectroscopy. Complexes formed in water at a w/w ratio of Si26H:DNA (pCMVbeta) of 300 were smallest with a mean hydrodynamic diameter of 83 nm. For effective condensation a w/w ratio of Si26H:DNA of 30 was sufficient. Further, the absorbed DNA was protected from enzymatic degradation by DNase I.
    背景与目标: :我们合成了具有共价键连接的阳离子表面修饰的二氧化硅纳米粒子(SiNP),并证明了它们具有静电结合,浓缩和保护质粒DNA的能力。这些颗粒可以用作基因载体的DNA载体。所有纳米颗粒的尺寸在10至100 nm之间,在pH 7.4下显示7至31 mV的表面电荷电位。它们是通过使用N-(2-氨基乙基)-3-氨基丙基三甲氧基硅烷或N-(6-氨基己基)-3-氨基丙基三甲氧基硅烷对市售(IPAST)或内部合成的二氧化硅颗粒进行改性制成的。所有颗粒均与pCMVbeta质粒DNA形成复合物,琼脂糖凝胶中DNA的比例依赖性阻滞作用以及可溶性DNA与纳米颗粒的共同沉淀作用证明了这一点。高盐和碱性pH确实抑制了复合物的形成。如通过光子相关光谱法所示,吸收到颗粒上也降低了质粒DNA的流体力学尺寸。在水中以Si26H:DNA(pCMVbeta)的重量比为300形成的复合物最小,平均流体动力学直径为83 nm。对于有效的缩合,Si26H:DNA的w / w比为30足够。此外,DNase I保护了吸收的DNA免受酶促降解。
  • 【X连锁隐性脊髓和延髓性肌萎缩的女性携带者的临床特征和偏斜的X染色体失活。】 复制标题 收藏 收藏
    DOI:10.1007/s004150170069 复制DOI
    作者列表:Ishihara H,Kanda F,Nishio H,Sumino K,Chihara K
    BACKGROUND & AIMS: :In X-linked recessive disorders, a few female gene carriers become symptomatic. Recent evidence implicates skewed X-chromosome inactivation in such female carriers. We studied the clinical features of eight female gene carriers of X-linked recessive spinal and bulbar muscular atrophy (SBMA), and evaluated the relationship between phenotype and genotype from the viewpoint of X-chromosome inactivation. Seven of eight cases were symptomatic, showing mild muscle weakness, frequent muscle cramps, slight elevation of the serum creatinine kinase level, or neurogenic changes on the electromyogram. Only one carrier was asymptomatic clinically. For the estimation of X-chromosome inactivation, the methylation status of the androgen receptor (AR) gene was determined by polymerase chain reaction-based assay. Highly skewed inactivation of the affected AR gene was found in the asymptomatic carrier, while symptomatic carriers had a random or lower inactivation pattern of the affected AR gene. These findings suggest that most female carriers of SBMA show some clinical abnormalities, and highly skewed inactivation of the affected X-chromosome seems to closely relate with escape of the manifestation in female carriers of SBMA.
    背景与目标: :在X连锁隐性疾病中,一些女性基因携带者有症状。最近的证据表明这种女性携带者中偏斜的X染色体失活。我们研究了X连锁隐性脊髓和延髓性肌萎缩症(SBMA)的八个女性基因携带者的临床特征,并从X染色体失活的角度评估了表型与基因型之间的关系。 8例中有7例是有症状的,表现出轻度的肌肉无力,频繁的肌肉痉挛,血清肌酐激酶水平的轻微升高或肌电图的神经源性改变。临床上仅一种携带者无症状。为了估计X染色体的失活,通过基于聚合酶链反应的测定来确定雄激素受体(AR)基因的甲基化状态。在无症状的携带者中发现受影响的AR基因高度失活,而有症状的携带者具有受影响的AR基因的随机或较低失活模式。这些发现表明,大多数SBMA女性携带者表现出某些临床异常,受影响的X染色体的高度偏斜失活似乎与SBMA女性携带者表现的逃避密切相关。
  • 【pH对脂双分子膜电导率的影响与由羧基载体grisorixin,alborixin和monensin诱导的碱性离子迁移有关。】 复制标题 收藏 收藏
    DOI:10.1016/0005-2736(78)90284-5 复制DOI
    作者列表:Sandeaux R,Seta P,Jeminet G,Alleaume M,Gavach C
    BACKGROUND & AIMS: :The influence of the pH on the stability and stoichiometry of the complexes formed by carboxylic-antibiotics such as grisorixin, alborixin and monensin with alkaline and alkaline earth cations has been investigated. The maximum values of bimolecular lipid membrane conductance are obtained with grisorixin and potassium ion. The conductance-pH curves show a very pronounced maximum in the neutral pH range. The results are analysed on the basis of a dimeric form of the ionophore in the complex and the possibility of having several charged complexes resulting from an heterogeneous reaction, the number of each complexed form depending on the pH of the bulk solutions.
    背景与目标: :已经研究了pH对由羧基抗生素(如格氏毒素,阿尔法毒素和莫能菌素)与碱金属和碱土金属阳离子形成的配合物的稳定性和化学计量的影响。用grisorixin和钾离子可获得双分子脂质膜电导的最大值。电导-pH曲线在中性pH范围内显示出非常明显的最大值。基于复合物中离子载体的二聚体形式以及由于异质反应而产生几种带电复合物的可能性来分析结果,每种复合形式的数量取决于本体溶液的pH。
  • 【核黄素载体在大鼠小肠和结肠中的转运功能:位点差异和三环型药物的作用。】 复制标题 收藏 收藏
    DOI:10.1080/107175401316906874 复制DOI
    作者列表:Tomei S,Yuasa H,Inoue K,Watanabe J
    BACKGROUND & AIMS: :The present study was aimed at kinetically characterizing the newly found carrier-mediated riboflavin transport system in the rat colon, comparing it with that in the small intestine, and also probing the potential roles of these transport systems in intestinal drug absorption. Riboflavin transport, evaluated by measuring the initial uptake into everted intestinal tissue sacs, was saturable with a Michaelis constant (Km) of 0.13 microM and a maximum transport rate (Jmax) of 0.74 pmol/min/100 mg wet tissue weight (wtw) in the colon. Both the Km and the Jmax were smaller than those (0.57 microM and 4.26 pmol/min/100 mg wtw, respectively) in the small intestine, suggesting that the transport system in the colon has a higher affinity to substrates and a smaller transport capacity than its counterpart in the small intestine. The carrier-mediated riboflavin transport in the colon, similarly to that in the small intestine, was Na+-dependent and inhibited by lumiflavin, a riboflavin analogue with an isoalloxazine ring, but not by D-ribose, which forms the side-chain attached to the isoalloxazine ring in riboflavin. To further clarify the substrate specificities of the transport systems, we examined the effects of several drugs with a tricyclic structure similar to isoalloxazine ring on riboflavin transport. Chlorpromazine, a phenothiazine derivative, was found to inhibit riboflavin transport in both the small intestine and the colon. Methylene blue also was found to be a potent inhibitor in both sites. These results suggest that some tricyclic-type drugs could interfere with intestinal riboflavin absorption by specific carrier-mediated transport systems. These transport systems may play roles in the absorption of tricyclic-type drugs.
    背景与目标: :本研究旨在动力学表征大鼠结肠中新发现的载体介导的核黄素转运系统,将其与小肠中的核黄素转运系统进行比较,并探讨这些转运系统在肠道药物吸收中的潜在作用。核黄素转运是通过测量肠外翻组织囊的初始吸收来评估的,其饱和度(米氏常数)(Km)为0.13 microM,最大转运速率(Jmax)为0.74 pmol / min / 100 mg湿组织重量(wtw)。冒号。 Km和Jmax均小于小肠中的Km和Jmax(分别为0.57 microM和4.26 pmol / min / 100 mg wtw),这表明结肠中的转运系统与底物的亲和力更高,转运能力比它在小肠中的对应物。载体介导的核黄素在结肠中的转运与小肠中的转运类似,是Na依赖性的,并受到lumiflavin(一种具有异四恶嗪环的核黄素类似物)的抑制,但不受D-核糖(形成侧链连接)的抑制。核黄素中的异恶嗪环。为了进一步阐明转运系统的底物特异性,我们检查了几种具有类似于异恶嗪环的三环结构的药物对核黄素转运的影响。已发现氯噻嗪(一种吩噻嗪衍生物)在小肠和结肠中均抑制核黄素转运。还发现亚甲基蓝在两个位点都是有效的抑制剂。这些结果表明,某些三环型药物可能会干扰特定载体介导的转运系统对肠道核黄素的吸收。这些转运系统可能在三环型药物的吸收中发挥作用。
  • 【用三苯基phosph官能化的半telechelic HPMA共聚物作为药物载体进行膜转导和线粒体定位。】 复制标题 收藏 收藏
    DOI:10.1021/bm060336m 复制DOI
    作者列表:Callahan J,Kopecek J
    BACKGROUND & AIMS: :Semitelechelic HPMA (N-(2-hydroxypropyl)methacrylamide) copolymers possessing a single terminal lipophilic triphenylphosphonium (TPP) cation and fluorescent labels were synthesized to determine how the attached cation affected cellular uptake and intracellular trafficking. In vitro mitochondrial uptake fluorescence quenching assays using isolated mouse liver mitochondria indicated that only lower molecular weight (<5 kDa) BODIPY FL-labeled TPP-semitelechelic HPMA copolymers exhibited significant organelle localization or uptake. In vitro cellular uptake and intracellular trafficking was evaluated using cultured human ovarian carcinoma cells. Cells incubated with all types of TPP copolymers used in the study appeared to internalize the polymer by endocytosis only, and all of the internalized copolymer was confined to the lysosomal compartment after 24 h. Endocytotic uptake of the TPP-HPMA copolymer conjugates was rapid, suggesting that they were internalized by adsorptive endocytosis, rather than fluid-phase pinocytosis. Low-molecular weight (<5 kDa) and high-molecular weight (>5 kDa) semitelechelic copolymers, microinjected into cultured cells indicated that the TPP moiety did not significantly localize the polymers to mitochondria.
    背景与目标: 合成具有单个末端亲脂性三苯基phosph(TPP)阳离子和荧光标记的半螯合HPMA(N-(2-羟丙基)甲基丙烯酰胺)共聚物,以确定连接的阳离子如何影响细胞摄取和细胞内运输。使用分离的小鼠肝线粒体进行的体外线粒体摄取荧光猝灭测定表明,只有较低分子量(<5 kDa)的BODIPY FL标记的TPP-半螯合HPMA共聚物显示出明显的细胞器定位或摄取。使用培养的人卵巢癌细胞评估了体外细胞摄取和细胞内运输。与研究中使用的所有类型的TPP共聚物一起孵育的细胞似乎仅通过内吞作用使聚合物内在化,并且所有内在化的共聚物在24小时后均被限制在溶酶体区室中。 TPP-HPMA共聚物共轭物的胞吞摄取很快,这表明它们是通过吸附性内吞作用而不是液相的胞吞作用而被内在化的。显微注射到培养细胞中的低分子量(<5 kDa)和高分子量(> 5 kDa)半遥望共聚物表明TPP部分并未将聚合物显着定位于线粒体。
  • 【α突触核蛋白基因中E46K突变的有症状和无症状携带者的心脏交感神经去神经。】 复制标题 收藏 收藏
    DOI:10.1016/j.parkreldis.2012.08.001 复制DOI
    作者列表:Tijero B,Gómez-Esteban JC,Lezcano E,Fernández-González C,Somme J,Llorens V,Martínez A,Ruiz-Martínez J,Foncea N,Escalza I,Berganzo K,Aniel-Quiroga MA,Ruiz V,Terán N,Kaufmann H,Zarranz JJ
    BACKGROUND & AIMS: OBJECTIVE:The aim of this study was to analyze autonomic function and cardiac sympathetic innervation in symptomatic and asymptomatic carriers of the E46K alpha-synuclein gene (SNCA) mutation. PATIENTS AND METHODS:Autonomic function tests were performed in six patients, four of whom were symptomatic carriers (ages: 46, 59, 52 and 28-years) and two who were asymptomatic carriers (ages: 52 and 29 years). Autopsy studies were performed on an additional two symptomatic carriers not eligible for autonomic testing. Patients completed the SCOPA autonomic questionnaire, and underwent the head-up tilt test accompanied by measurements of plasma norepinephrine. Valsalva maneuver and deep breathing tests, along with recording of sympathetic skin response (SSR) and cardiac MIBG scintigraphy were carried out. Myocardial tissue sections removed from the two autopsied cases were subjected to routine histological staining and immunohistochemical processing with monoclonal antibodies against tyrosine hydroxylase and alpha-synuclein. RESULTS:Both the four symptomatic and the older asymptomatic carriers reported abnormalities in the SCOPA questionnaire and had markedly diminished cardiac MIBG uptake. Plasma norepinephrine in the supine and tilted positions was normal in all subjects. Only one patient had significant orthostatic hypotension. There was a complete absence of tyrosine hydroxylase immunostaining in the myocardium of the two autopsied cases. INTERPRETATION:We have found imaging and histological evidence of cardiac sympathetic denervation in symptomatic and asymptomatic carriers of the E46K alpha-synuclein gene mutation. The sympathetic denervation appears to be organ-specific, with selective affectation of the heart given that plasma norepinephrine levels and blood pressure were normal.
    背景与目标: 目的:本研究旨在分析E46Kα-突触核蛋白基因(SNCA)突变的有症状和无症状携带者的自主神经功能和心脏交感神经。
    患者和方法:对六名患者进行了自主功能检查,其中四名是有症状的携带者(年龄:46、59、52和28岁),两名是无症状的携带者(年龄:52和29岁)。对另外两个不适合进行自主性检查的症状携带者进行了尸检研究。患者完成了SCOPA自主调查表,并进行了抬头倾斜测试,并同时测量了血浆去甲肾上腺素。进行了Valsalva动作和深呼吸测试,并记录了交感性皮肤反应(SSR)和心脏MIBG闪烁显像。从两个尸检病例中取出的心肌组织切片用酪氨酸羟化酶和α-突触核蛋白的单克隆抗体进行常规组织学染色和免疫组织化学处理。
    结果:这四个有症状的和较年长的无症状携带者均在SCOPA调查表中报告异常,并且心脏MIBG摄取明显减少。在所有受试者中,仰卧位和倾斜位的血浆去甲肾上腺素均正常。只有一名患者患有明显的体位性低血压。在两个尸检病例的心肌中完全没有酪氨酸羟化酶免疫染色。
    解释:我们在E46Kα-突触核蛋白基因突变的有症状和无症状携带者中发现了心脏交感神经支配的影像学和组织学证据。鉴于血浆去甲肾上腺素水平和血压正常,交感神经似乎是器官特异性的,对心脏有选择性的影响。
  • 【隐性严重1型和3型von Willebrand病(VWD),无症状杂合子携带者与血型O相关的von Willebrand因子缺乏症和主要1型VWD的特征。】 复制标题 收藏 收藏
    DOI:10.1177/1076029606291401 复制DOI
    作者列表:Michiels JJ,Berneman Z,Gadisseur A,van der Planken M,Schroyens W,van de Velde A,van Vliet H
    BACKGROUND & AIMS: :Recessive type 3 von Willebrand disease (VWD) is caused by homozygosity or double heterozygosity for two non-sense mutations (null alleles). Type 3 VWD is easy to diagnose by the combination of a strongly prolonged bleeding time (BT), absence of ristocetine-induced platelet aggregation (RIPA), absence of von Willebrand factor (VWF) protein, and prolonged activated partial thromboplastin time (aPTT) due to factor VIII:coagulant (FVIII:C) deficiency. VWD type 3 is associated with a pronounced tendency to mucocutaneous and musculoskeletal bleedings since early childhood. Carriers of one null allele are usually asymptomatic at VWF levels of 50% of normal. Recessive severe type 1 VWD is caused by homozygosity or double heterozygosity for a missense mutation. Recessive type 1 VWD differs from type 3 VWD by the presence of detectable von Willebrand factor: antigen VWF:Ag and FVIII:C levels between 0.09 and 0.40 U/mL. Patients with recessive type 1 VWD show an abnormal VWF multimeric pattern in plasma and/or platelets consistent with severe type 2 VWD. Carriers of a missense mutation may have mild bleeding and mild VWF deficiency and can be diagnosed by a double VWF peak on cross immunoelectrophoresis (CIE). There will be cases of mild and moderate recessive type 1 VWD due to double heterozygosity of two missense mutations, or with the combination of one missense mutation with a non-sense or bloodgroup O. Mild deficiency of VWF in the range of 0.20 to 0.60 U/mL, with normal ratios of von Willebrand factor: ristocetine cofactor/antigen VWF:RCo/Ag and VWF:collagen binding/antigen (VWF:CB/Ag), normal VWF multimers, and a completely normal response to desmopressin acetate (DDAVP) with VWF level rising from below to above 1.00 U/mL are very likely cases of so-called pseudo-VWF deficiency in individuals with normal VWF protein and gene. Autosomal dominant type 1 VWD variants are in fact type 2 variants caused by a heterozygous missense mutation in the VWF gene that produces a mutant VWF protein that has a dominant effect on normal VWF protein produced by the normal VWF allele with regard to the synthesis, processing, storage, secretion, and/or proteolysis of VWF in endothelial cells. A DDAVP challenge test clearly differentiates between dominant type 1 VWD phenotype and dominant type 2 M VWD.
    背景与目标: :隐性3型von Willebrand病(VWD)是由两个无义突变的纯合子或双重杂合子(无效等位基因)引起的。通过强烈延长出血时间(BT),不存在瑞斯托汀诱导的血小板聚集(RIPA),不存在von Willebrand因子(VWF)蛋白和延长活化部分凝血活酶时间(aPTT)的组合,很容易诊断3型VWD由于VIII因子:凝血剂(FVIII:C)缺乏。自幼儿期以来,VWD 3型与明显的粘膜皮肤和肌肉骨骼出血倾向有关。一个无效等位基因的携带者在VWF正常水平的50%时通常是无症状的。隐性严重1型VWD是由错义突变的纯合或双重杂合引起的。隐性1型VWD与3型VWD的区别在于存在可检测的von Willebrand因子:抗原VWF:Ag和FVIII:C水平在0.09至0.40 U / mL之间。隐性1型VWD患者在血浆和/或血小板中显示出异常的VWF多聚体模式,与严重的2型VWD一致。错义突变的携带者可能有轻度出血和轻度VWF缺乏症,可以通过交叉免疫电泳(CIE)上的双VWF峰进行诊断。由于两个错义突变的双重杂合性,或者一个错义突变与无义或血型O的组合,将出现轻度和中度隐性1型VWD。VWF的轻度缺乏在0.20至0.60 U的范围内/ mL,具有正常比例的von Willebrand因子:ristocetine辅因子/抗原VWF:RCo / Ag和VWF:胶原结合/抗原(VWF:CB / Ag),正常的VWF多聚体,以及对醋酸去氨加压素(DDAVP)的反应完全正常VWF水平从低于1.00 U / mL升至1.00 U / mL以上的人,很有可能在具有正常VWF蛋白和基因的个体中发生所谓的伪VWF缺乏症。常染色体显性1型显性VWD变体实际上是2型变异,由VWF基因中的杂合错义突变引起,该突变产生突变的VWF蛋白,在合成,加工方面对正常VWF等位基因产生的正常VWF蛋白具有显性作用内皮细胞中VWF的储存,分泌,和/或蛋白水解。 DDAVP挑战测试清楚地区分了显性1型VWD表型和显性2 M VWD型。
  • 【包含微球作为载体的可生物降解的壳聚糖支架,可用于软骨组织工程的受控转化生长因子β1递送。】 复制标题 收藏 收藏
    DOI: 复制DOI
    作者列表:Cai DZ,Zeng C,Quan DP,Bu LS,Wang K,Lu HD,Li XF
    BACKGROUND & AIMS: BACKGROUND:Natural articular cartilage has a limited capacity for spontaneous regeneration. Controlled release of transforming growth factor-beta1 (TGF-beta1) to cartilage defects can enhance chondrogenesis. In this study, we assessed the feasibility of using biodegradable chitosan microspheres as carriers for controlled TGF-beta1 delivery and the effect of released TGF-beta1 on the chondrogenic potential of chondrocytes. METHODS:Chitosan scaffolds and chitosan microspheres loaded with TGF-beta1 were prepared by the freeze-drying and the emulsion-crosslinking method respectively. In vitro drug release kinetics, as measured by enzyme-linked immunosorbent assay, was monitored for 7 days. Lysozyme degradation was performed for 4 weeks to detect in vitro degradability of the scaffolds and the microspheres. Rabbit chondrocytes were seeded on the scaffolds containing TGF-beta1 microspheres and incubated in vitro for 3 weeks. Histological examination and type II collagen immunohistochemical staining was performed to evaluate the effects of released TGF-beta1 on cell adhesivity, proliferation and synthesis of the extracellular matrix. RESULTS:TGF-beta1 was encapsulated into chitosan microspheres and the encapsulation efficiency of TGF-beta1 was high (90.1%). During 4 weeks of incubation in lysozyme solution for in vitro degradation, the mass of both the scaffolds and the microspheres decreased continuously and significant morphological changes was noticed. From the release experiments, it was found that TGF-beta1 could be released from the microspheres in a multiphasic fashion including an initial burst phase, a slow linear release phase and a plateau phase. The release amount of TGF-beta1 was 37.4%, 50.7%, 61.3%, and 63.5% for 1, 3, 5, and 7 days respectively. At 21 days after cultivation, type II collagen immunohistochemical staining was performed. The mean percentage of positive cells for collagen type II in control group (32.7% +/- 10.4%) was significantly lower than that in the controlled TGF-beta1 release group (92.4% +/- 4.8%, P < 0.05). Both the proliferation rate and production of collagen type II in the transforming growth factor-beta1 microsphere incorporated scaffolds were significantly higher than those in the scaffolds without microspheres, indicating that the activity of TGF-beta1 was retained during microsphere fabrication and after growth factor release. CONCLUSION:Chitosan microspheres can serve as delivery vehicles for controlled release of TGF-beta1, and the released growth factor can augment chondrocytes proliferation and synthesis of extracellular matrix. Chitosan scaffolds incorporated with chitosan microspheres loaded with TGF-beta1 possess a promising potential to be applied for controlled cytokine delivery and cartilage tissue engineering.
    背景与目标: 背景:天然关节软骨的自发再生能力有限。转化生长因子-beta1(TGF-beta1)向软骨缺损的控制释放可以增强软骨生成。在这项研究中,我们评估了使用可生物降解的壳聚糖微球作为载体控制TGF-β1传递的可行性以及释放的TGF-β1对软骨细胞软骨形成潜能的影响。
    方法:采用冻干法和乳液交联法分别制备载有TGF-β1的壳聚糖支架和壳聚糖微球。通过酶联免疫吸附测定法测量的体外药物释放动力学被监测7天。进行溶菌酶降解4周以检测支架和微球的体外降解性。将兔软骨细胞接种在含有TGF-β1微球的支架上,并在体外孵育3周。进行组织学检查和II型胶原免疫组织化学染色,以评估释放的TGF-β1对细胞粘附性,细胞外基质增殖和合成的影响。
    结果:TGF-β1被包埋在壳聚糖微球中,TGF-β1的包封率较高(90.1%)。在溶菌酶溶液中温育4周以进行体外降解的过程中,支架和微球的质量持续下降,并且形态发生了显着变化。从释放实验中,发现TGF-β1可以以多相方式从微球释放,包括初始爆发阶段,缓慢的线性释放阶段和平稳阶段。 TGF-beta1在1、3、5和7天的释放量分别为37.4%,50.7%,61.3%和63.5%。培养后21天,进行II型胶原免疫组织化学染色。对照组中II型胶原蛋白阳性细胞的平均百分比(32.7%/-10.4%)显着低于对照TGF-beta1释放组(92.4%/-4.8%,P <0.05)。掺入了转化生长因子-β1微球的支架中的II型胶原的增殖速率和产生均显着高于不含微球的支架中的II型胶原的生长,这表明TGF-β1的活性在微球制备过程中和生长因子释放后得以保留。
    结论:壳聚糖微球可以作为TGF-β1控释的载体,释放的生长因子可以促进软骨细胞的增殖和细胞外基质的合成。壳聚糖支架与负载TGF-β1的壳聚糖微球相结合,具有可用于控制细胞因子递送和软骨组织工程的潜力。
  • 【作为口服药物递送载体的复合水凝胶中胰岛素的体外释放行为和稳定性。】 复制标题 收藏 收藏
    DOI:10.1016/s0378-5173(03)00378-8 复制DOI
    作者列表:Kim B,Peppas NA
    BACKGROUND & AIMS: :Novel pH-responsive complexation hydrogels containing pendent glucose (P(MAA-co-MEG)) or grafted PEG chains (P(MAA-g-EG)) were synthesized by photopolymerization. The feasibility of these hydrogels as oral protein delivery carriers was evaluated. The pH-responsive release behavior of insulin was analyzed from both P(MAA-co-MEG) and P(MAA-g-EG) hydrogels. In acidic media (pH 2.2), insulin release from the hydrogels was very slow. However, as the pH of the medium was changed to 6.5, a rapid release of insulin occurred. In both cases, the biological activity of insulin was retained. For P(MAA-co-MEG) hydrogels, the biological activity of insulin decreased when the pendent glucose content increased. In P(MAA-g-EG) hydrogels, when the grafted PEG molecular weight increased, the insulin biological activity decreased. Finally, hydrogels of P(MAA-co-MEG) prepared with an initial ratio of 1:4 MEG:MAA and P(MAA-g-EG) hydrogels containing PEG chains of molecular weights of 200 showed the greatest change in insulin release rate from acidic to basic pH solutions and the greatest protective effect for insulin in simulated GI tract conditions.
    背景与目标: :通过光聚合反应合成了含有悬垂葡萄糖(P(MAA-co-MEG))或接枝PEG链(P(MAA-g-EG))的新型pH响应络合水凝胶。评价了这些水凝胶作为口服蛋白质递送载体的可行性。从P(MAA-co-MEG)和P(MAA-g-EG)水凝胶中分析了胰岛素的pH响应释放行为。在酸性介质(pH 2.2)中,胰岛素从水凝胶中的释放非常缓慢。但是,随着培养基的pH值更改为6.5,胰岛素迅速释放。在这两种情况下,胰岛素的生物活性均得以保留。对于P(MAA-co-MEG)水凝胶,当垂下葡萄糖含量增加时,胰岛素的生物活性降低。在P(MAA-g-EG)水凝胶中,当嫁接的PEG分子量增加时,胰岛素的生物活性降低。最后,初始比例为1:4 MEG:MAA的P(MAA-co-MEG)水凝胶和分子量为200的PEG链的P(MAA-g-EG)水凝胶显示最大的胰岛素释放速率变化从酸性到碱性pH溶液,在模拟胃肠道条件下对胰岛素的保护作用最大。

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