The first Chinese case of Creutzfeldt-Jakob disease patient with R208H mutation in PRNP

A case of Creutzfeldt-Jakob disease (CJD) with a rare mutation of the prion protein (PrP) gene (PRNP) at codon 208 (R208H), while the codon 129 was a methionine homozygous genotype is reported. The patient initial displayed hand tremor, dizziness and progressive cognitive dysfunction. Subsequently, other symptoms gradually appeared, including cerebellar ataxia and mental disorder. No periodic activity was recorded at electroencephalography (EEG) and 14-3-3 protein in cerebrospinal fluid was negative. Total clinical course was about four months. Retrospective investigation of this family across seven generations did not figure out clear family history. However, genetic analyses revealed six first-degree family members with the R208H allele.Key words: creutzfeldt-Jakob disease, PRNP, R208H     

Integrated dataset of screening hits against multiple neglected disease pathogens

New chemical entities are desperately needed that overcome the limitations of existing drugs for neglected diseases. Screening a diverse library of 10,000 drug-like compounds against 7 neglected disease pathogens resulted in an integrated dataset of 744 hits. We discuss the prioritization of these hits for each pathogen and the strong correlation observed between compounds active against more than two pathogens and mammalian cell toxicity. Our work suggests that the efficiency of early drug discovery for neglected diseases can be enhanced through a collaborative, multi-pathogen approach.     

Tet1 is dispensable for maintaining pluripotency and its loss is compatible with embryonic and postnatal development

The Tet family of enzymes (Tet1/2/3) converts 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC). Mouse embryonic stem cells (mESCs) highly express Tet1 and have an elevated level of 5hmC. Tet1 has been implicated in ESC maintenance and lineage specification in?vitro but its precise function in development is not well defined. To establish the role of Tet1 in pluripotency and development, we have generated Tet1 mutant mESCs and mice. Tet1(-/-) ESCs have reduced levels of 5hmC and subtle changes in global gene expression, and are pluripotent and support development of live-born mice in tetraploid complementation assay, but display skewed differentiation toward trophectoderm in?vitro. Tet1 mutant mice are viable, fertile, and grossly normal, though some mutant mice have a slightly smaller body size at birth. Our data suggest that Tet1 loss leading to a partial reduction in 5hmC levels does not affect pluripotency in ESCs and is compatible with embryonic and postnatal development.     

Infection of H69AR cells with retroviral particles harboring interfering RNAi significantly reduced the multidrug resistance of these small cell lung cancer cells

Incubation of the drug-sensitive H69, a small cell lung cancer cell line, with increased concentrations of adriamycin yielded multidrug resistant (MDR) H69AR cells that over-express multidrug resistance-associated protein (MRP1). MRP1 co-transports its substrate with glutathione (GSH), leading to lower intracellular GSH. In this report we tested whether depleting intracellular GSH in MRP1-expressing cells could hyper-sensitize them to anticancer drugs or not. We have found that the GSH contents in MRP1-expressing cells are significantly lower than their corresponding control cells. The treatment with MRP1 substrate verapamil or the GSH synthetase inhibitor buthionine sulfoxi-mine significantly reduced the intracellular GSH contents in MRP1-expressing cells. Interestingly, depleting intracellular GSH contents can hyper-sensitize the MRP1-cDNA transfected BHK cells to daunomycin, but not the adriamycin-selected H69AR cells. Further analyses indicated that anti-apoptotic factor Bcl2 might be a factor responsible for the fact that depleting intracellular GSH could not hyper-sensitize H69AR cells to daunomycin. We hypothesized that knocking down the expression of Bcl2 could hyper-sensitize H69AR cells to daunomycin. Interestingly, infection of H69AR cells with retroviral particles harboring Bcl2 interfering RNAi not only reduced the expression of Bcl2, but also many factors that contribute to MDR, such as Bcl-xl, MRP1 and ABCC3, etc., leading to the MDR H69AR cells more sensitive to daunomycin than the parental H69 cell. Thus, although the mechanisms of the down-regulation of the genes contributing to MDR remain to be elucidated, retroviral particles harboring Bcl2 interfering RNAi could be used as an alternative way to sensitize the MDR cancer cells to anticancer drugs.     

广谱抗真菌转基因水稻植株化学成分与结构动态变化

【背景】种植广谱抗真菌水稻可能会带来一定的环境生物安全问题,对其植株的化学成分进行实质等同性分析是转基因水稻安全性评价的重要内容之一。【方法】以表达广谱抗真菌蛋白转基因水稻转品1和转品8及其相应非转基因水稻七丝软粘的秸秆为研究材料,采用化学法和扫描电镜技术分析外源基因的导入对水稻秸秆化学成分以及组织显微结构的影响。【结果】（1）在整个生长发育过程中,广谱抗真菌转基因水稻转品1和转品8与其非转基因水稻七丝软粘叶片、叶鞘和茎的纤维素、半纤维素、木质素以及粗蛋白含量的变化趋势基本一致,且品种间化学成分的含量不存在显著差异。（2）广谱抗真菌转基因水稻叶片表皮的硅质瘤状结构以及气孔的形状和致密程度与其非转基因水稻七丝软粘相似;茎壁、厚壁组织、薄壁组织以及大小维管束的形态和分布情况未发生明显变化。【结论与意义】表达广谱抗真菌蛋白转基因水稻秸秆的化学成分和组织显微结构与非转基因水稻基本一致。这为广谱抗真菌转基因水稻的环境安全性评估提供了依据。     

Leptospira interrogans encodes an ROK family glucokinase involved in a cryptic glucose utilization pathway

Although Leptospira interrogans is unable to utilize glucose as its carbon/energy source, the LA_1437 gene of L. interrogans serovar Lai potentially encodes a group III glucokinase (GLK). The L. interrogans GLK (LiGLK) heterogeneously expressed in Escherichia coli was purified and proved to be a homodimeric enzyme with its specific activity of 12.3 ± 0.6 U/mg x protein determined under an improved assay condition (pH 9.0, 50 ° C), 7.5-fold higher than that assayed under the previously used condition (pH 7.3, 25 ° C). The improved sensitivity allowed us to detect this enzymatic activity of (5.0 ± 0.6) × 10(-3) U/mg x protein in the crude extract of L. interrogans serovar Lai cultured in standard Ellinghausen-McCullough-Johnson-Harris medium. The k(cat) and K(m) values for d-glucose and ATP were similar to those of other group III GLKs, although the K(m) value for ATP was slightly higher. Site-directed mutagenesis analysis targeting the conserved amino acid residues in the potential ATP-binding motif hinted that a proper array of Gly residues in the motif might be important for maintaining the conformation that was essential for its function. Gene expression profiling and quantitative proteomic data mining provided preliminary evidence for the absence of efficient systems involved in glucose transport and glycolysis that might account for the failure of glucose utilization in L. interrogans.     

光强在低温弱光胁迫后番茄叶片光合作用恢复中的作用

为了研究光强在低温弱光胁迫后番茄叶片光合作用恢复中的作用,以番茄品种浙粉202为材料,研究了低温弱光后恢复期全光照与遮荫对光合作用和叶绿素荧光参数的影响。结果表明：低温弱光（8℃/12℃,PFD 80 μmol·m^-2·s^-1）导致番茄叶片P_n、Φ_PSⅡ、qP和F_v′/F_m′的下降,但诱导了NPQ的上升,未引起F_v/F_m的变化;全光照（100%光照）下恢复1 使得植株叶片P_n、F_v/F_m、Φ_PSⅡ、qP、NPQ和F_v′/F_m′均大幅下降,随后光合和荧光参数可缓慢恢复至对照水平;遮荫（40%光照）恢复植株F_v/F_m、Φ_PSⅡ和F_v′/F_m′仅在第一天稍有下降,而P_n和qP还略有上升,NPQ虽有所降低但仍显著高于对照水平,随后光合和荧光参数均可迅速恢复到对照水平。说明低温弱光虽抑制了光合作用的进行,但并未引起光抑制的发生;全光照恢复加剧了叶片光抑制的发生,而遮荫恢复可通过叶片PSⅡ光化学活性的快速恢复和天线色素热耗散能力的增强以保护光合机构免受伤害,有利于光合作用的迅速恢复。     

明日叶查尔酮对小鼠肝癌细胞PCNA 和BCL-2 蛋白表达影响

目的:研究明日叶(Angelica keiskei Koidz)查尔酮对小鼠肝癌细胞PCNA和BCL-2蛋白表达的影响。方法:将50只皮下接种肝癌H22细胞株的小鼠随机分为5组,每组10只。高、中、低剂量组分别每日经口灌胃给予40、20、5mg/kg的查尔酮,肿瘤对照组给予等量生理盐水,连续10d,环磷酰胺组隔天腹腔注射环磷酰胺20mg/kg。取肝癌组织用四甲基偶氮噻唑蓝(MTT)法测各组小鼠肝癌细胞增殖活性,免疫组化法检测各组肝癌细胞增殖细胞核抗原(PCNA)和凋亡相关蛋白BCL-2表达水平。结果:高剂量查尔酮组和肿瘤对照组的肝癌细胞增殖活性分别为(0.716±0.018)和(1.135±0.032),差别有显著性(P<0.05)。高剂量组PCNA和BCL-2蛋白表达率分别为28.33%和16.77%,肿瘤对照组分别为72.77%和65.17%,差异均有显著性(P<0.05)。结论:查尔酮可降低小鼠肝癌细胞PCNA和BCL-2表达水平,对肝癌细胞增殖有一定抑制作用。     

大黄素对HT-29 细胞MAPKs 信号通路活化及IL-8 分泌的影响

目的:研究大黄素对IFN-和LPS刺激的人结肠癌细胞株HT-29细胞的ERK、JNK和p38 MARK和IL-8表达的影响。方法:人结肠癌细胞株HT-29细胞与40 ng/mL的IFN-共培养12 h,再加入100 ng/mL LPS刺激15 min,用大黄素预处理进行干预。ELISA检测HT-29细胞内的ERK、JNK和p38 MARK含量和细胞上清IL-8含量。结果:IFN-γ和LPS刺激后HT-29细胞的ERK、JNK和p38 MARK磷酸化水平和IL-8分泌明显升高。大黄素对p38和JNK磷酸化有明显的抑制作用,而对ERK磷酸化则没有明显抑制作用;大黄素能显著降低IFN-γ+LPS所引起的HT-29细胞IL-8的大量产生,并且呈明显的剂量依赖关系。结论:大黄素能有效抑制IFN-γ+LPS所引起的HT-29细胞p38和JNK的磷酸化,并显著降低IL-8分泌。