Fasting up‐regulates ferroportin 1 expression via a Ghrelin/GHSR/MAPK signaling pathway

The significant positive correlation between ghrelin and iron and hepcidin levels in the plasma of children with iron deficiency anemia prompted us to hypothesize that ghrelin may affect iron metabolism. Here, we investigated the effects of fasting or ghrelin on the expression of hepcidin, ferroportin 1 (Fpn1), transferrin receptor 1 (TfR1), ferritin light chain (Ft‐L) proteins, and ghrelin, and also hormone secretagogue receptor 1 alpha (GHSR1α) and ghrelin O‐acyltransferase (GOAT) mRNAs in the spleen and/or macrophage. We demonstrated that fasting induces a significant increase in the expression of ghrelin, GHSR1α, GOAT, and hepcidin mRNAs, as well as Ft‐L and Fpn1 but not TfR1 proteins in the spleens of mice in vivo. Similar to the effects of fasting on the spleen, ghrelin induced a significant increase in the expression of Ft‐L and Fpn1 but not TfR1 proteins in macrophages in vitro. In addition, ghrelin was found to induce a significant enhancement in phosphorylation of ERK as well as translocation of pERK from the cytosol to nuclei. Furthermore, the increased pERK and Fpn1 induced by ghrelin was demonstrated to be preventable by pre‐treatment with either GHSR1α antagonist or pERK inhibitor. Our findings support the hypothesis that fasting upregulates Fpn1 expression, probably via a ghrelin/GHSR/MAPK signaling pathway.     

盐土和沙土对新疆常见一年生盐生植物生长和体内矿质组成的影响

选用古尔班通古特沙漠南缘荒漠-绿洲交错带常见的一年生盐生植物盐角草、刺毛碱蓬、叉毛蓬、猪毛菜和碱地肤为材料,比较了它们在原状盐土和沙土中的生长及体内矿质元素组成的差异。结果表明：① 原状盐土0—100 cm各土层的pH值低于沙土,但电导率和含水量明显高于沙土;② 原状盐土中生长的植物干重是沙土中生长植物干重的7—118倍,后者的根冠比是前者的2—6倍。③ 体现肉质化程度的地上部含水率为52%—81%,中低耐盐植物含水率在两种土壤中差异显著,强耐盐植物差异不显著;④ 5种一年生盐生植物地上部氮浓度为11—34 g/kg,在有效氮含量高的盐土上植物氮浓度也高;磷浓度为1—4 g/kg,在有效磷高的盐土上植物磷浓度也较高（盐角草除外）;但沙土中的植物地上部钾浓度明显高于盐土中植株的地上部钾浓度,这与两种土壤在0—60 cm土层中的钾浓度差异相反;⑤ 尽管原状盐土0—100 cm土层中的水溶性钙、镁、钠、氯、硫浓度显著高于沙土,盐土与沙土中生长的植物地上部钠、水溶性氯和硫的浓度比值远远低于土壤中的相应元素浓度的比值,甚至盐土中的植株钙、镁浓度等于或显著低于沙土中生长的植物。表明盐土不仅影响一年生盐生植物的生长,也显著影响这些植物对矿质元素的吸收和累积。一年生盐生植物能够选择性吸收不同生境中的矿质元素。本研究期望为进一步深入研究盐生植物耐盐的适应机制提供依据,也可为植物修复盐碱土的品种选择提供参考。     

大肠杆菌甘油激酶基因(glpK)和甘油脱氢酶基因(gldA)的敲除及其对甘油产量的影响

利用Red重组系统构建了大肠杆菌JM109甘油激酶基因(glpK)和甘油脱氢酶基因(gldA)缺失的双突变菌株JM109B,然后将表达酿酒酵母3-磷酸甘油脱氢酶基因(GPD1)和3-磷酸甘油酯酶基因(HOR2)的质粒pSE-gpd1-hor2转化到JM109B突变菌株中,在含1%葡萄糖的摇瓶发酵培养基中37℃发酵24 h,甘油的最高产量为5.61 g/L,是原始菌株JM109/pSE-gpd1-hor2甘油产量的1.59倍;在30 L发酵罐中发酵28 h,甘油的最高产量为103.12 g/L,是原始菌株JM109/pSE-gpd1-hor2甘油产量的1.59倍,是原始菌株BL21/pSE-gpd1-hor2甘油产量的1.41倍,葡萄糖转化率为50.39%。     

青冈常绿阔叶林内的小气候特征

分析我国中亚热带东部青冈（Ｑｕｅｒｃｕｓｇｌａｕｃａ）常绿阔叶林内的小气候特征,１９９３～１９９５年的研究结果表明：①到达青冈林的总太阳辐射为３３４４７８０ｋＪ／（ｍ２·ａ）,四季中群落的反射率、透射率和吸收率分别为１６％～２２％、９％～１２％和６７％～７４％。②林冠外上方及群落上层气温在白天高于群落下层,夜间低于群落下层,可相差３～５℃,夏季差异最大。③林内外空气相对湿度的日动态呈“Ｕ”型变化,林内夜间湿度高达９０％左右,午间较低,在５０％左右。在四季的晴天中,林冠上方的空气相对湿度均低于林内,相差５％～２２％,夏季和冬季差异最大。④林中的ＣＯ２浓度在林冠层最低,近地面层最高,各季节始终低于林外,其中夏、秋两季最明显。⑤在春、夏、秋３季中,土壤温度为白天高于夜间,而冬季则为夜间高于白天;土壤湿度以冬、春季较高（３１．９％和２８．５％）,夏季最低（１４．２％）。由于青冈次生林的叶面积指数较小,群落结构较简单,因而整个群落的透光系数较大,群落内外空气温湿度的差异也较小,体现出幼年林向中年林过渡阶段的特点     

裂解性复制诱导产生可视化重组Epstein Barr病毒

为了在病毒的整个基因组中研究基因的功能,分析基因与基因之间的相互作用,含有整个野生型EB病毒(EBV)基因组的BAC-EBV质粒(p2089),首先被转染EBV阴性的HEK293细胞,经潮霉素筛选建立了HEK293/p2089稳定细胞系.再构建pcDNA3.1( )/BZLF1和pcDNA3.1( )/BALF4真核表达质粒,共转染至HEK293/p2089细胞内,诱导EBV裂解性复制产生可视化的重组EBV颗粒.重组EBV颗粒感染Raji细胞,在倒置荧光显微镜下和流式细胞仪记数GFP阳性细胞,根据这些"绿色Raji单位"确定病毒的滴度.在国内首次建立这种以细菌人工染色体(BAC)为基础的EBV感染性克隆技术,将允许对EB病毒基因组中任何基因的任何遗传修饰,为在整个基因组中对EB病毒基因功能的研究奠定了基础,也为对EBV与其相关的肿瘤如鼻咽癌发生机理的研究建立了新的技术平台.     

Rapid and efficient site-directed mutagenesis by single-tube 'megaprimer' PCR method.

We describe a rapid and efficient megaprimer PCR procedure for site-directed mutagenesis that does not require any intermediate purification of DNA between the two rounds of PCR. This protocol is based on the design of forward and reverse flanking primers with significantly different melting temperatures ( T m). A megaprimer is synthesized in the first PCR reaction using a mutagenic primer, the low T m flanking primer and a low annealing temperature. The second PCR reaction is performed in the same tube as the first PCR and utilizes the high T m flanking primer, the megaprimer product of the first PCR and a high annealing temperature, which prevents priming by the low T m primer from the first PCR reaction. We have used this protocol with two different plasmids to produce cDNAs encoding seven distinct mutated proteins. We have observed an average mutagenesis efficiency of 82% in these experiments.     

Allele-specific targeting of hsa-miR-657 to human IGF2R creates a potential mechanism underlying the association of ACAA-insertion/deletion polymorphism with type 2 diabetes

The biological mechanism of a recent discovered association of type 2 diabetes with the ACAA-insertion/deletion polymorphism at the 3′UTR of the IGF2R gene has remained unclear. A very recently emerging novel polymorphic control layer by microRNAs (miRNAs) makes it possible to elucidate this issue. In this study, a prediction by web tools MicroInspector and miRanda demonstrated that DNA sequence polymorphism (DSPs) ACAA-insertion/deletion in IGF2R 3′UTR is located within the hsa-miR-657 and hsa-miR-453 binding sites. And luciferase reporter assay revealed that hsa-miR-657 acts directly at the 3′UTR of the IGF2R. Furthermore, ACAA-deletion exerted a further repression compared with ACAA-insertion, indicating that hsa-miR-657 regulates IGF2R gene expression in a polymorphic control manner. Importantly, we also demonstrated that hsa-miR-657 can translationally regulate the IGF2R expression levels in Hep G2 cells. Thus, our findings testify the possibility that the ACAA-insertion/deletion polymorphism may result in the change of IGF2R expression levels at least in part by hsa-miR-657-mediated regulation, contributing to the elucidation for the pathogenesis of type 2 diabetes and raise the possibility that miRNAs or in combination with functional DNA sequence polymorphism may be valuable in the treatment of human type 2 diabetes.     

Autophagy: a novel guardian of HCV against innate immune response

Autophagy is an evolutionarily conserved process that catabolizes intracellular components and maintains cellular homeostasis. Autophagy involves the sequestration of cytoplasmic content within a double-membraned autophagosome, and the fusion of the autophagosome with a lysosome to form an autolysosome for subsequent degradation (Fig. 1A). Autophagy plays a pivotal role in various aspects of cellular responses to stresses, such as nutrient deprivation, damaged organelles, aggregated proteins, exposure to endoplasmic reticulum (ER) stress and pathogen infections. Virus infection often leads to ER stress and induction of the unfolded protein response (UPR). Recent studies reveal that virus-induced UPR may activate autophagy to support the virus life cycle. However, the exact roles of the UPR and autophagy in host cell-virus interactions are still enigmatic.     

慢病毒载体法制备红色荧光蛋白转基因小鼠

目的利用慢病毒介导的转基因方法制备红色荧光蛋白（mRFP）转基因小鼠,并建立转基因小鼠的技术平台。方法将携带mRFP基因的慢病毒注入ICR鼠单细胞受精卵卵周隙以感染受精卵,胚胎移植进假孕母鼠以获得仔代鼠,然后应用小动物活体成像仪、体视荧光显微镜和PCR等鉴定并获得mRFP转基因鼠。结果移植卵周隙注射有慢病毒的胚胎40枚给2只假孕母鼠,共获得仔鼠6只;利用体视荧光显微镜检测mRFP表达,在蛋白水平证实6只F0代中,2只（R3和R4）鼠耳高表达mRFP,其余的弱表达mRFP（R1、R2和R5）或荧光强度（R6）与野生型ICR鼠无明显差别,而DNA水平检测证实,6只F0代中,5只（R1、R2、R3、R4和R5）基因组中整合有外源转基因hUb-mRFP,预示基因型鉴定结果很好验证了体视荧光显微镜鉴定结果。此外,mRFP转基因首建鼠基因组中整合的mRFP基因可稳定遗传和表达。结论建立了慢病毒法快速制备转基因小鼠的技术平台,这为针对不同基因建立相应转基因小鼠以实现恒定或条件性的转基因过表达或RNA干涉（RNAi）,并进而在体内解析相应基因功能和建立人类疾病模型等奠定了坚实基础。     

生物质热裂解生产生物燃料的研究进展

生物质能源是一种绿色的可以替代化石能源的一种可再生的能源。尽管高温分解生物质处于发展阶段,但在目前水平,高温裂解因其可以在氧存在下热分解将生物材料直接转化为固态,液态和气态能源产品而受到广泛关注。本文介绍了生物质的热裂解,包括慢速热裂解、快速热裂解、闪解、催化热裂解等过程,重点讨论了在各种生物质材料的热裂解过程中各种操作参数如温度和生物粒子大小等对生物燃料收率的影响。