中国流行性乙型脑炎病毒表型和基因型的研究进展

流行性乙型脑炎(乙脑)病毒是引起病毒性脑炎最主要的病毒之一,本文对我国分离株的表型和基因型特性进行了综合分析。生物学特征研究显示,不同毒株间存在空斑形态、小鼠神经侵入致病性、保护性抗原和血凝性的明显差异。中国在1977年前,自然界中仅存在基因Ⅲ型乙脑病毒,但自1977年以后基因Ⅰ型和Ⅲ型病毒均从自然界分离到,而基因Ⅰ型病毒已成为优势病毒,其中多数分离自蚊虫。基因序列分析显示,两种基因型的氨基酸序列差异率很小(≤3%),与当前广泛应用的减毒活疫苗株比较,仅≤3%的氨基酸序列差异率,这些差别主要存在于与SA14-14-2减毒相关的位点,提示SA14-14-2疫苗能够有效地保护这两种基因型毒株的感染。     

Enhanced phenol bioavailability by means of photocatalysis

Phenol was investigated for the ability of TiO₂ photocatalysis to increase its bioavailability as an electron donor for denitrification. The rate of nitrate removal by denitrification was increased by up to 2.6-fold by exposing phenol to photocatalysis for 30 min, although the rate decreased with increasing photocatalysis. The increased denitrification rate appeared to be associated with the photocatalytic production of carboxylic acids, but the slow down correlated to the production of catechol and hydroquinone.     

Occurrence of 4-tert-butylphenol (4-t-BP) biodegradation in an aquatic sample caused by the presence of Spirodela polyrrhiza and isolation of a 4-t-BP-utilizing bacterium

Although 4-tert-butylphenol (4-t-BP) is a serious aquatic pollutant, its biodegradation in aquatic environments has not been well documented. In this study, 4-t-BP was obviously and repeatedly removed from water from four different environments in the presence of Spirodela polyrrhiza, giant duckweed, but 4-t-BP persisted in the environmental waters in the absence of S. polyrrhiza. Also, 4-t-BP was not removed from autoclaved pond water with sterilized S. polyrrhiza. These results suggest that the 4-t-BP removal from the environmental waters was caused by biodegradation stimulated by the presence of S. polyrrhiza rather than by uptake by the plant. Moreover, Sphingobium fuliginis OMI capable of utilizing 4-t-BP as a sole carbon and energy source was isolated from the S. polyrrhiza rhizosphere. Strain OMI degraded 4-t-BP via a meta-cleavage pathway, and also degraded a broad range of alkylphenols with linear or branched alkyl side chains containing two to nine carbon atoms. Root exudates of S. polyrrhiza stimulated 4-t-BP degradation and cell growth of strain OMI. Thus, the stimulating effects of S. polyrrhiza root exudates on 4-t-BP-degrading bacteria might have contributed to 4-t-BP removal in the environmental waters with S. polyrrhiza. These results demonstrate that the S. polyrrhiza–bacteria association may be applicable to the removal of highly persistent 4-t-BP from wastewaters or polluted aquatic environments.     

Pro-inflammatory cytokines modulate iron regulatory protein 1 expression and iron transportation through reactive oxygen/nitrogen species production in ventral mesencephalic neurons

Both inflammatory processes associated with microglia activation and abnormal iron deposit in dopaminergic neurons are involved in the pathogenesis of Parkinson's disease (PD). However, the relationship between neuroinflammation and iron accumulation was not fully elucidated. In the present study, we aimed to investigate whether the pro-inflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) released by microglia, could affect cellular iron transportation in primary cultured ventral mesencephalic (VM) neurons. The results showed that IL-1β or TNF-α treatment led to increased ferrous iron influx and decreased iron efflux in these cells, due to the upregulation of divalent metal transporter 1 with the iron response element (DMT1 + IRE) and downregulation of ferroportin1 (FPN1). Increased levels of iron regulatory protein 1 (IRP1), transferrin receptor 1 (TfR1) and hepcidin were also observed in IL-1β or TNF-α treated VM neurons. IRP1 upregulation could be fully abolished by co-administration of radical scavenger N-acetyl-l-cysteine and inducible NO synthetase inhibitor N_ω-nitro-l-arginine methyl ester hydrochloride. Further experiments demonstrated that IL-1β and TNF-α release was remarkably enhanced by iron load in activated microglia triggered by lipopolysaccharide or 1-methyl-4-phenylpyridinium (MPP⁺). In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-intoxicated mice, salicylate application could not block DMT1 + IRE upregulation in dopaminergic neurons of substantia nigra. These results suggested that IL-1β and TNF-α released by microglia, especially under the condition of iron load, might contribute to iron accumulation in VM neurons by upregulating IRP1 and hepcidin levels through reactive oxygen/nitrogen species production. This might provide a new insight into unraveling that microglia might aggravate this iron mediated neuropathologies in PD.     

衡阳紫色土丘陵坡地不同恢复阶段土壤基础呼吸及代谢熵的变化

采用空间序列代替时间序列的方法,对衡阳紫色土丘陵坡地不同过程中的不同土层、根际(Rhizosphere,R)与非根际(Non-rhizosphere,S)的土壤基础呼吸(Soil basal respiration,SBR)及代谢熵(Metabolic quotient,qCO2)的变化特征以及它们与土壤理化性质的关系进行研究。结果表明：(1)不同恢复阶段SBR与qCO2存在明显差异,从裸地(Ⅰ)、草本群落(Ⅱ)、灌木群落(Ⅲ)至乔木群落阶段(Ⅳ),SBR显著增强,qCO2显著减小,反映土地质量正在逐渐恢复;(2)不同恢复阶段SBR与qCO2存在垂直变化特征,从0～20cm、20～40cm至40～60cm土层, SBR显著减弱,qCO2显著增加;(3)从Ⅱ→Ⅳ,R与S变化明显,R与S的SBR均显著增强, R/S显著减小,表现出R＞S的特点,R与S的qCO2的逐渐减小, R/S逐渐上升,也表现出R＞S的特点,因此,创造更好的土壤条件有利于该区域植被恢复。     

Three‐dimensional neural differentiation of embryonic stem cells with ACM induction in microfibrous matrices in bioreactors

The clinical use of pluripotent stem cell (PSC)‐derived neural cells requires an efficient differentiation process for mass production in a bioreactor. Toward this goal, neural differentiation of murine embryonic stem cells (ESCs) in three‐dimensional (3D) polyethylene terephthalate microfibrous matrices was investigated in this study. To streamline the process and provide a platform for process integration, the neural differentiation of ESCs was induced with astrocyte‐conditioned medium without the formation of embryoid bodies, starting from undifferentiated ESC aggregates expanded in a suspension bioreactor. The 3D neural differentiation was able to generate a complex neural network in the matrices. When compared to 2D differentiation, 3D differentiation in microfibrous matrices resulted in a higher percentage of nestin‐positive cells (68% vs. 54%) and upregulated gene expressions of nestin, Nurr1, and tyrosine hydroxylase. High purity of neural differentiation in 3D microfibrous matrix was also demonstrated in a spinner bioreactor with 74% nestin + cells. This study demonstrated the feasibility of a scalable process based on 3D differentiation in microfibrous matrices for the production of ESC‐derived neural cells. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1013–1022, 2013     

Atomic force microscopy imaging of live mammalian cells

Atomic force microscopy (AFM) was used to examine the morphology of live mammalian adherent and suspended cells. Time-lapse AFM was used to record the locomotion dynamics of MCF-7 and Neuro-2a cells. When a MCF-7 cell retracted, many small sawtooth-like filopodia formed and reorganized, and the thickness of cellular lamellipodium increased as the retraction progressed. In elongated Neuro-2a cells, the cytoskeleton reorganized from an irregular to a parallel, linear morphology. Suspended mammalian cells were immobilized by method combining polydimethylsiloxane-fabricated wells with poly-L-lysine electrostatic adsorption. In this way, the morphology of a single live lymphoma cell was imaged by AFM. The experimental results can improve our understanding of cell locomotion and may lead to improved immobilization strategies.     

A novel strategy to inhibit the reproduction and translation of hepatitis C virus

Hepatitis C virus (HCV), a positive single-stranded RNA virus, is a major cause of liver disease in humans. Herein we report a novel strategy to inhibit the reproduction and translation of HCV using a short RNA, named an Additional RNA, to activate the endonuclease activity of Argonaute 2 (Ago2). In the presence of the Additional RNA, the HCV genome RNA has the requisite 12 nucleotides of base-pairing with microRNA-122. This activates the endonuclease activity of Ago2, resulting in cleavage and release of the HCV genome RNA from Ago2 and microRNA-122. The free HCV genome RNA would be susceptible to intracellular degradation, effectively inhibiting its reproduction and translation. This study presents a new method to inhibit HCV that may hold great potential for HCV treatment in the future.