• 【BRCA1和BRCA2种系突变携带者的预防标本中的偶然癌,重点是输卵管病变:6例病例报告并复习文献。】 复制标题 收藏 收藏
    DOI:10.1097/01.pas.0000202161.80739.ac 复制DOI
    作者列表:Carcangiu ML,Peissel B,Pasini B,Spatti G,Radice P,Manoukian S
    BACKGROUND & AIMS: :The identification of germ-line mutations in 2 genes (BRCA1 and BRCA2) responsible for the majority of hereditary ovarian cancers has led an increasing number of women carriers of these mutations to undergo prophylactic oophorectomy (PO) to reduce their risk of subsequent ovarian carcinoma. A large number of unexpected, clinically occult neoplasms are thus being discovered. Up to December 2004, the Medical Genetics Service of the National Cancer Institute in Milan, Italy, has tested 756 probands from breast and/or ovarian cancer families for BRCA1 and BRCA2 germ-line mutations. Molecular screening of family members led to the identification of 344 female carriers of BRCA1 (239) or BRCA2 (105) germ-line mutations. Of the 186 potentially eligible women (37 of whom had tested positive for BRCA1 and 13 for BRCA2 mutation), 50 (26.8%) chose to undergo PO. Six clinically occult primary gynecologic malignancies (2 stage IIIC serous carcinomas of the ovary, 3 in situ serous carcinomas of the fallopian tube, and 1 stage IIB invasive serous carcinoma of the fallopian tube) and 1 occult ovarian metastasis from breast carcinoma were identified in the PO specimens of 7 women (all BRCA1 mutated). Four of the patients with occult primary gynecologic cancers are alive without disease 129, 87, 38, and 7 months after PO, respectively. One of the 2 patients with primary ovarian cancer and the single patient with tubal invasive carcinoma are alive with recurrent disease 83 and 20 months after PO, respectively. In addition, one of the patients whose PO specimen did not show any malignancy presented with stage IIIC tubal carcinoma 77 months after PO. The relatively high number of tubal neoplasms found at PO in this group of patients underlines the linkage between mutation and the risk of developing tubal cancer, and stresses the need to include removal of the entire tubes at the time of PO and of thoroughly evaluating the specimens at the microscopic level. The upstaging of all 3 invasive carcinomas after staging surgery, and the late recurrence and persistence of 2 of them despite treatment indicate that small size of the tumors should not preclude therapy.
    背景与目标: :对负责大多数遗传性卵巢癌的2个基因(BRCA1和BRCA2)的种系突变的鉴定已导致越来越多的女性携带这些突变的女性进行预防性卵巢切除术(PO),以降低其患上卵巢癌的风险。因此,发现了大量意想不到的临床隐匿性肿瘤。截至2004年12月,意大利米兰国家癌症研究所的医学遗传学服务已对756个来自乳腺癌和/或卵巢癌家族的先证者进行了BRCA1和BRCA2种系突变测试。家庭成员的分子筛查导致识别出BRCA1(239)或BRCA2(105)种系突变的344个雌性携带者。在186名可能合格的女性中(其中37名对BRCA1测试呈阳性,13名BRCA2突变检测为阳性),其中50名(26.8%)选择接受PO。在临床中发现了6例临床隐匿的原发性妇科恶性肿瘤(2例IIIC期卵巢浆液性癌,3例输卵管原位浆液性癌和1例IIB期输卵管浸润性浆液性癌)和1例隐匿性卵巢癌卵巢转移。 7名妇女的PO标本(所有BRCA1突变)。分别在PO后129、87、38和7个月,有四名患有隐匿性原发性妇科癌症的患者还活着而没有疾病。 2例原发性卵巢癌患者中的1例和输卵管浸润性癌的单例患者在PO后分别存活83个月和20个月。此外,其中一名PO标本未显示任何恶性肿瘤的患者在PO后77个月出现IIIC期输卵管癌。该组患者在PO中发现的相对较多的输卵管肿瘤强调了突变与发生输卵管癌的风险之间的联系,并强调需要在PO时切除整个管并彻底评估标本在微观层面上。分期手术后所有3种浸润性癌的分期升级,并且尽管有治疗,但其中2种仍较晚复发和持续存在,这表明较小的肿瘤不应排除治疗的可能性。
  • 【航空保险的重要因素和风险管理策略:对台湾航空公司的实证研究。】 复制标题 收藏 收藏
    DOI:10.1111/j.1539-6924.2008.01036.x 复制DOI
    作者列表:Lin YH,Chang YH
    BACKGROUND & AIMS: :Aviation insurance premiums have become a heavy burden for the airline industry since September 11, 2001. Although the industry must constantly balance its operations between profitability and safety, the reality is that airlines are in the business of making money. Therefore, their ability to reduce cost and manage risk is a key factor for success. Unlike past research, which used subjective judgment methods, this study applied quantitative historical data (1999-2000) and gray relation analysis to identify the primary factors influencing ratemaking for aviation insurance premiums. An empirical study of six airlines in Taiwan was conducted to determine these factors and to analyze the management strategies used to deal with them. Results showed that the loss experience and performance of individual airlines were the key elements associated with aviation insurance premiums paid by each airline. By identifying and understanding the primary factors influencing ratemaking for aviation insurance, airlines will better understand their relative operational strengths and weaknesses, and further help top management identify areas for further improvement. Knowledge of these factors combined with effective risk management strategies, may result in lower premiums and operating costs for airline companies.
    背景与目标: 自2001年9月11日以来,航空保险费已成为航空业的沉重负担。尽管航空业必须在盈利能力和安全性之间不断地平衡其运营,但现实是航空公司从事的是赚钱业务。因此,他们降低成本和管理风险的能力是成功的关键因素。与以往使用主观判断方法的研究不同,本研究使用定量历史数据(1999-2000年)和灰色关联分析来确定影响航空保险费率制定的主要因素。对台湾六家航空公司进行了实证研究,以确定这些因素并分析了用于处理这些因素的管理策略。结果表明,单个航空公司的损失经历和业绩是与每个航空公司支付的航空保险费相关的关键因素。通过识别和理解影响航空保险费率制定的主要因素,航空公司将更好地了解其相对的经营优势和劣势,并进一步帮助高层管理人员确定需要进一步改进的领域。了解这些因素并结合有效的风险管理策略,可能会降低航空公司的保费和运营成本。
  • 【携带9号染色体的家族相互平衡易位的携带者的生殖随访,并与预期结果进行比较。】 复制标题 收藏 收藏
    DOI: 复制DOI
    作者列表:Castorina P,Rodeschini O,Nocera G,Larizza L
    BACKGROUND & AIMS: :Reproductive follow-up of carriers of familial reciprocal balanced translocations involving chromosome 9 and comparison with predicted outcome: Chromosome 9 is commonly implicated in reciprocal translocations (rcp). Twenty-seven families segregating rcp involving chromosome 9 were selected with the aim of comparing the theoretical risk of Mental Retardation with Congenital Anomalies (MCA/MR) calculated according to Human Cytogenetics Forum with the observed reproductive follow-up. The 27 families include 157 subjects. The reproductive follow-up showed that the majority of mothers underwent full-term pregnancies (88/130), and that there were 37 spontaneous and five voluntary abortions. Eighty-one subjects were karyotyped: 18 had a normal karyotype, 50 carried an rcp, ten had an unbalanced rcp-related karyotype and three an abnormal rcp-unrelated karyotype. Of the 88 live-born individuals, seven had an abnormal rcp-related karyotype with partial chromosome 9 trisomy (four cases) or partial 9p monosomy (three cases), and 48 were rcp carriers, two of whom also presented additional anomalies. The evaluation of reproductive outcomes in the 27 families studied revealed good concordance between the Human Cytogenetics Forum predictions and the observed follow-up in relation to the most probable mode of unbalance at birth, and the higher risk of MCA/MR in rcp carriers with unbalanced live-borns in comparison with those generating healthy progeny
    背景与目标: :涉及染色体9的家族相互平衡易位的携带者的生殖随访,并与预期结果进行比较:染色体9通常与相互易位(rcp)有关。选择了27个涉及9号染色体的rcp分离家庭,目的是将根据人类细胞遗传学论坛计算的先天性精神发育迟滞的理论风险与观察到的生殖随访进行比较。 27个家庭包括157个科目。生殖随访表明,大多数母亲都进行了足月妊娠(88/130),并且有37例自然流产和5例自愿流产。八十一名受试者进行了核型分析:18名受试者具有正常的核型,五十名受试者具有rcp,十名受试者具有不平衡的rcp相关性核型,三名受试者具有异常的rcp无关性核型。在88个活产个体中,有7个具有与rcp相关的染色体核型异常,其中9号染色体三体性(4例)或9p单体性染色体3个(3例),而rcp携带者48个,其中两个还存在其他异常。对所研究的27个家庭的生殖结果进行的评估表明,人类细胞遗传学论坛的预测与观察到的关于出生时最可能的失衡方式以及在失衡的rcp携带者中MCA / MR的风险较高之间的随访之间保持了良好的一致性。与产生健康后代的活产婴儿相比
  • 【基质囊泡是骨形态发生蛋白(BMP),血管内皮生长因子(VEGF)和非胶原基质蛋白的载体。】 复制标题 收藏 收藏
    DOI:10.1007/s00774-008-0859-z 复制DOI
    作者列表:Nahar NN,Missana LR,Garimella R,Tague SE,Anderson HC
    BACKGROUND & AIMS: :Matrix vesicles (MVs) are well positioned in the growth plate to serve as a carrier of morphogenetic information to nearby chondrocytes and osteoblasts. Bone morphogenetic proteins (BMPs) carried in MVs could promote differentiation of these skeletal cells. Vascular endothelial growth factor (VEGF) in MVs could stimulate angiogenesis. Therefore, a study was undertaken to confirm the presence of bone morphogenetic protein (BMP)-1 through-7, VEGF, and the noncollagenous matrix proteins, bone sialoprotein (BSP), osteopontin (OPN), osteocalcin (OC), and osteonectin (ON) in isolated rat growth plate MVs. MVs were isolated from collagenase-digested rachitic rat tibial and femoral growth plates. The presence of BMP-1 through BMP-7, VEGF, BSP, ON, OPN, and OC was evaluated by Western blot, plus ELISA analyses for BMP-2 and-4 content. The alkaline phosphatase-raising ability of MV extracts on cultured rat growth plate chondrocytes was measured as a reflection of MV ability to promote chondroosseous differentiation. BMP-1 through-7, VEGF, BSP, ON, OPN, and OC were all detected by Western blot analyses. Chondrocytes treated with MV extracts showed a two-to threefold increase in alkaline phosphatase activity over control, indicating increased differentiation. Significant amounts of BMP-2 and BMP-4 were detected in MVs by ELISA. Combined, these data suggest that MVs could play an important morphogenetic role in growth plate and endochondral bone formation.
    背景与目标: 基质小泡(MVs)很好地位于生长板上,可作为形态学信息的载体,传递给附近的软骨细胞和成骨细胞。 MV中携带的骨形态发生蛋白(BMP)可以促进这些骨骼细胞的分化。 MV中的血管内皮生长因子(VEGF)可以刺激血管生成。因此,进行了一项研究以确认存在骨形态发生蛋白(BMP)-1至7,VEGF和非胶原基质蛋白,骨唾液蛋白(BSP),骨桥蛋白(OPN),骨钙蛋白(OC)和骨粘连蛋白( ON)在分离的大鼠生长板MV中。从胶原酶消化的大鼠胫骨和股骨生长板中分离出MV。通过蛋白质印迹以及ELISA分析BMP-2和-4的含量,评估BMP-1到BMP-7,VEGF,BSP,ON,OPN和OC的存在。测量MV提取物在培养的大鼠生长板软骨细胞上的碱性磷酸酶升高能力,反映了MV促进软骨骨分化的能力。通过蛋白质印迹分析检测到BMP-1至7,VEGF,BSP,ON,OPN和OC。用MV提取物处理的软骨细胞显示碱性磷酸酶活性比对照提高了2到3倍,表明分化增加。通过ELISA检测到MV中有大量的BMP-2和BMP-4。综合起来,这些数据表明,MVs在生长板和软骨内骨形成中可以发挥重要的形态发生作用。
  • 【壳聚糖改性的透明质酸基纳米药物载体。】 复制标题 收藏 收藏
    DOI:10.1016/j.ijbiomac.2020.01.118 复制DOI
    作者列表:Turcsányi Á,Varga N,Csapó E
    BACKGROUND & AIMS: :Fabrication possibilities, detailed size and structural characterization of biodegradable chitosan (Chit) polysaccharide-modified hyaluronic acid (HyA)-based colloidal carriers are demonstrated. The negatively charged and highly hydrophilic HyA polymer chains have been ionically modified by positively charged pure Chit and crosslinked Chit macromolecules at various Chit/HyA weight ratios, which resulted in the formation of carrier nanoparticles (NPs) having three different nanostructures depending on the polymer concentrations. Electrostatically-compensated Chit/HyA polymer coils with loose colloidal structure, tripolyphosphate (TPP)-crosslinked Chit-TPP/HyA NPs having interpenetrating polymer network and well-defined Chit-TPPcore-HyAshell NPs with diameters of 100-300 nm were also prepared and were loaded with tocopherol (TCP) and cholecalciferol (D3) having Vitamin E and D activity, respectively. By using rheological, particle charge titration and conductivity studies we first confirmed that the expected 1:1 Chit/HyA monomer molar ratio is strongly influenced by the pH of the polymer solutions as well as the deacetylation degree of Chit which are crucial factors for the solubility, purity and the quality of the commercially available biocompatible Chit in aqueous medium. Encapsulation studies revealed that D3 could be better incorporated in every system, especially in Chit-TPP/HyA NPs, while for TCP the simple Chit/HyA polymer coils were the most promising carriers.
    背景与目标: :展示了可生物降解的壳聚糖(Chit)多糖修饰的透明质酸(HyA)基胶体载体的制备可能性,详细的尺寸和结构特征。带负电荷且亲水性高的HyA聚合物链已通过带正电荷的纯Chit和交联的Chit高分子在各种Chit / HyA重量比下进行了离子修饰,这导致形成了根据聚合物浓度具有三种不同纳米结构的载体纳米颗粒(NPs) 。还制备了具有胶体结构松散的静电补偿Chit / HyA聚合物线圈,具有互穿聚合物网络的三聚磷酸盐(TPP)交联的Chit-TPP / HyA NP和直径100-300nm的轮廓分明的Chit-TPPcore-HyAshell NP。分别装载具有维生素E和D活性的生育酚(TCP)和胆钙化固醇(D3)。通过流变,颗粒电荷滴定和电导率研究,我们首先证实预期的1:1 Chit / HyA单体摩尔比受聚合物溶液的pH值和Chit脱乙酰度的强烈影响,这是溶解度的关键因素水性介质中市售的生物相容性Chit的纯度,质量和质量。封装研究表明,D3可以更好地集成到每个系统中,尤其是在Chit-TPP / HyA NP中,而对于TCP,简单的Chit / HyA聚合物线圈是最有前途的载体。
  • 【Caco-2细胞单层中锰转运的动力学和锰转运载体的基因表达。】 复制标题 收藏 收藏
    DOI:10.1007/s10534-013-9670-y 复制DOI
    作者列表:Li X,Xie J,Lu L,Zhang L,Zhang L,Zou Y,Wang Q,Luo X,Li S
    BACKGROUND & AIMS: :Two experiments were conducted to investigate the kinetics of manganese (Mn) transport in Caco-2 cell monolayers and the gene expressions of Mn transport carriers in apical (AP) and basolateral (BL) membranes. In experiment 1, the cells were treated with the medium containing 146 μmol/L of Mn (MnSO4·H2O). Both the uptake and transport of Mn from AP-BL or from BL-AP at different time-points were assessed to determine the optimal time for kinetics of Mn transport. The transport of Mn increased linearly with higher efficiency values in AP-BL than in BL-AP direction, however, the uptake of Mn revealed an asymptotic pattern within 120 min. In experiment 2, the kinetics of Mn transport in AP-BL was determined with media containing Mn concentrations from 0 to 2,500 μmol/L at 40 and 120 min, respectively, and mRNA levels of divalent metal transporter 1 (DMT1) and ferroportin (FPN1) were determined in Caco-2 cells treated with the medium containing 0 or 800 μmol/L of Mn for 120 min. The kinetics of Mn transport showed a carrier-mediated process when Mn concentrations were lower than 1,000 μmol/L and a linear increment when Mn concentrations exceeded 1,000 μmol/L at either 40 or 120 min. Mn treatment decreased (P < 0.01) DMT1 mRNA level and increased (P < 0.01) FPN1 mRNA level. The results from the present study suggested that Mn transport in AP-BL fit both carrier-mediated saturable and non-saturable diffusion processes, and Mn transport carriers DMT1 and FPN1 mediate the apical uptake and basolateral exit of Mn in Caco-2 cells.
    背景与目标: :进行了两个实验,以研究锰(Mn)在Caco-2细胞单层中的迁移动力学以及Mn(转运)载体在顶(AP)和基底外侧(BL)膜中的基因表达。在实验1中,用含146μmol/ L Mn(MnSO4·H2O)的培养基处理细胞。评估了在不同时间点从AP-BL或BL-AP吸收和转运Mn的情况,以确定Mn转运动力学的最佳时间。在AP-BL中,Mn的运输以比BL-AP方向高的效率值线性增加,但是,Mn的吸收显示120分钟内的渐近模式。在实验2中,使用含有Mn浓度从0到2500μmol/ L的培养基分别在40和120分钟测定AP-BL中Mn转运的动力学,以及二价金属转运蛋白1(DMT1)和铁转运蛋白(FPN1)的mRNA水平。在用含0或800μmol/ L Mn的培养基处理120分钟的Caco-2细胞中测定)。当Mn浓度低于1,000μmol/ L时,Mn迁移的动力学表现出一个由载体介导的过程,而当Mn浓度超过1,000μmol/ L时,在40或120 min时,其迁移呈线性增加。锰处理降低(P <0.01)DMT1 mRNA水平和增加(P <0.01)FPN1 mRNA水平。本研究的结果表明,AP-BL中的Mn转运既符合载体介导的饱和和非饱和扩散过程,而Mn转运载体DMT1和FPN1介导Caco-2细胞中Mn的根尖吸收和基底外侧出口。
  • 【如何管理高风险无症状COVID-19携带者; COVID-19对公共卫生和皮肤病学专业也构成了巨大威胁。】 复制标题 收藏 收藏
    DOI:10.1111/dth.14064 复制DOI
    作者列表:Kassir M,Gupta M,Abdelmaksoud A,Goldust M
    BACKGROUND & AIMS: -2
    背景与目标: -2
  • 8 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。

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