Experimental chronic hepatitis B infection of neonatal tree shrews (Tupaia belangeri chinensis): A model to study molecular causes for susceptibility and disease progression to chronic hepatitis in humans

ABSTRACT: BACKGROUND: Hepatitis B virus (HBV) infection continues to be an escalating global health problem. Feasible and effective animal models for HBV infection are the prerequisite for developing novel therapies for this disease. The tree shrew (Tupaia) is a small animal species evolutionary closely related to humans, and thus is permissive to certain human viral pathogens. Whether tree shrews could be chronically infected with HBV in vivo has been controversial for decades. Most published research has been reported on adult tree shrews, and only small numbers of HBV infected newborn tree shrews had been observed over short time periods. We investigated susceptibility of newborn tree shrews to experimental HBV infection as well as viral clearance over a protracted time period. RESULTS: Forty-six newborn tree shrews were inoculated with the sera from HBV-infected patients or tree shrews. Serum and liver samples of the inoculated animals were periodically collected and analyzed using fluorescence quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, Southern blot, and immunohistochemistry. Six tree shrews were confirmed and four were suspected as chronically HBV-infected for more than 48 (up to 228) weeks after inoculation, including three that had been inoculated with serum from a confirmed HBV-infected tree shrew. CONCLUSIONS: Outbred neonatal tree shrews can be long-term chronically infected with HBV at a frequency comparable to humans. The model resembles human disease where also a smaller proportion of infected individuals develop chronic HBV related disease. This model might enable genetic and immunologic investigations which would allow determination of underlying molecular causes favoring susceptibility for chronic HBV infection and disease establishment vs. viral clearance.     

桂林喀斯特地区克隆生长红背山麻杆种群的点格局分析

植物空间分布格局是种群自身特性、种间关系以及环境条件综合作用的结果。对植物空间格局和关联性的研究有助于认识植物群落的形成、维持和演替过程。红背山麻杆(Alchornea trewioides(Benth.)Muell.Arg.)是桂林喀斯特地区重要的先锋物种。调查了桂林喀斯特地区以克隆繁殖为主的红背山麻杆群落,采用点格局方法,以Ripley's K函数为基础运用g(r)和L(r)函数对红背山麻杆种群进行点格局以及不同径级之间关联性分析。结果表明:(1)红背山麻杆在小尺度(小于2 m)上表现为聚集分布;在大尺度(大于2 m)上表现为随机分布;(2)3个径级之间在小尺度两两表现为正关联。随着尺度的增大,3径级之间的关联性由正关联转变为无关联或者负关联;(3)在大尺度范围红背山麻杆较大地径的母株对其他母株的克隆小分株产生较强的抑制作用,随着两个径级大小差异增大,它们的空间关联性会逐渐呈无关联或负关联,与径级Ⅰ与径级Ⅱ及径级Ⅱ与径级Ⅲ相比,径级Ⅰ与径级Ⅲ的空间关联性呈较强的负关联。     

代谢工程中基因修饰与基因表达的工具

代谢工程利用重组DNA技术导入定向改造的基因 ,以改进微生物细胞的某些代谢特性 ,已经发展成为一个工业微生物育种和优化发酵过程的强有力工具。基因的修饰与表达是代谢工程的重要组成部分。本文介绍了近年来代谢工程中基因修饰与表达所用的工具方面的进展。     

Hypoxia-induced brain cell damage in male albino wistar rat

The biochemical markers of rat under low oxygen concentration, including brain water level, lactic acid, necrosis and Na+-K+-ATPase, was detected to analyze the hypoxia-induced brain damage, and to analyze the mechanism of brain injury. Histopathological alteration in brain tissue induced by hypoxia were investigated through hematoxylin and eosin stain (HE). Hypoxia induced factor-1a (HIF-1a) expression level in the brain was carried out using immunohistochemistry. Lactic acid level was positively correlated with the level of hypoxia, while concentration-dependent decrease in total Na+-K+-ATPase activity was noted. Hypoxia induced rathad a significant difference on brain water content compared to controls. The level of necrosis and lactic acid level was increased, and the decrease of Na+-K+-ATPase activity was observed. Histopathological examination of brain confirmed that there was neuronal cell death in hippocampal region. HIF-1a expression increased the hypoxia adaptation capability of the rat through the expressions of genes. Lactic acid, Na+-K+-ATPase and HIF-1a plays an important role in brain injury.     

Decreased autophagy was implicated in the decreased apoptosis during decidualization in early pregnant mice

Folate deficiency is a major risk factor of birth defects. Mechanistic studies on folate deficiency resulting in birth defects have mainly focused on fetal development. There have been few studies on folate deficiency from the point of view of the mother’s uterus. In our previous study, we demonstrated that folate deficiency inhibits apoptosis of decidual cells, thereby restraining decidualization of the endometrium and impairing pregnancy. In this study, we further investigated the potential mechanism by which folate deficiency decreases endometrial apoptosis during decidualization. To investigate whether endometrium autophagy was inhibited under folate deficiency during decidualization, we performed real-time PCR for endometrial LC3 and P62 on day 6 (D6) to D8 of pregnancy in mice, and both were significantly changed compared to non-folate-deficient mice. Western blots showed that LC3-II and P62 were also changed in folate-deficient mice. Compared with control mice, a few punctuate LC3-II structures were detected in the folate deficiency group by immunofluorescence. Transmission electron micrographs of decidual cells on D8 showed that there were no evident autophagosomes in the folate deficiency group. In addition, apoptosis-related protein analysis by western blotting, TUNEL staining and flow cytometry showed that decreased endometrial apoptosis on D8 of pregnancy under folate deficiency was reversed after treatment with rapamycin, an autophagy inducer. ROS measurement showed that the endometrium ROS level was reduced by folate deficiency and that rapamycin reversed this effect on day 8 of pregnancy. All the results suggest that inhibiting endometrial autophagy may be implicated in the decreased endometrial apoptosis under folate deficiency during decidualization.     

Chondrogenic differentiation of human mesenchymal stem cells: a comparison between micromass and pellet culture systems

High-density cell culture is pivotal for the chondrogenic differentiation of human mesenchymal stem cells (hMSCs). Two high-density cell culture systems, micromass and pellet culture, have been used to induce chondrogenic differentiation of hMSCs. In micromass culture, the induced-cartilage tissues were larger, more homogenous and enriched in cartilage-specific collagen II but the fibrocartilage-like feature, collagen I, and hypertrophic chondrocyte feature, collagen X, were markedly decreased compared to those in pellet culture. Furthermore, real time RT-PCR analysis demonstrated that collagen II and aggrecan mRNA were up-regulated while collagen X and collagen I mRNA were down-regulated in micromass culture. Thus, the micromass culture system is a promising tool for in vitro chondrogenic studies.     

COP1 Jointly Modulates Cytoskeletal Processes and Electrophysiological Responses Required for Stomatal Closure

Reorganization of the cortical microtubule cytoskeleton is critical for guard cell function. Here, we investigate how environmental and hormonal signals cause these rearrangements and find that COP1, a RING-finger-type ubiquitin E3 ligase, is required for degradation of tubulin, likely by the 26S proteasome. This degradation is required for stomatal closing. In addition to regulating the cytoskeleton, we show that cop1 mutation impaired the activity of S-type anion channels, which are critical for stomatal closure. Thus, COP1 is revealed as a potential coordinator of cytoskeletal and electrophysiological activities required for guard cell function.     

Hydroxyl-functionalized DNAs with different linkers and their complementary duplex stability

Tert-butyldiphenylsilyl (TBDPS) was testified to be an appropriate orthogonal protecting group for novel 7-hydroxyl-functionalized 8-aza-7-deaza-2'-deoxyadenosine analogues. It was stable in partial and complete hydrogenation reactions used for the different linker preparation. The corresponding phosphoramidites and hydroxyl-functionalized oligodeoxynucleotides were synthesized and identified. The thermal effect of the hydroxyl group with different linkers on DNA duplexes was evaluated. It provided a feasible strategy for the preparation of hydroxyl-functionalized DNAs for the nucleic acid research.     

A non-viral vector for potential DMD gene therapy study by targeting a minidystrophin-GFP fusion gene into the hrDNA locus

Gene therapy has emerged as a promising approach for the lethal disorder of Duchenne muscular dystrophy （DMD）. Using a novel non-viral delivery system, the human ribosomal DNA （hrDNA） targeting vector, we tar- geted a minidystrophin-GFP fusion gene into the hrDNA locus of HT1080 cells with a high site-specific integrated efficiency of 10-s, in which the transgene could express efficiently and continuously. The minidystrophin-GFP fusion protein was easily found to localize on the plasma membrane of HT1080 cells, indicating its possible physiologic performance. Our findings showed that the hrDNAtargeting vector might be highly useful for DMD gene therapy study.     

老龙皮总氨基酸的提取工艺研究

通过单因素选择和正交设计法,以老龙皮总氨基酸含量为指标,分别对乙醇浓度、料液比、提取温度、提取时间等因素进行考查,优选老龙皮总氨基酸的最佳提取条件。结果显示料液比对总氨基酸提取率影响最大,最佳提取条件为60%乙醇作提取液,料液比(W/V)为1∶20,在70℃提取3 h。