Abstract:Silencing target genes with small regulatory RNAs is widely used to investigate gene function and therapeutic drug development. Recently,triplex-based approaches have provided another attractive means to achieve targeted gene regulation and gene manipulation at the molecular and cellular levels. Nuclear entry of oligonucleotides and enhancement of their af?nity to the DNA targets are key points of such approaches. In this study, we developed lipid-based transport of a locked-nucleic-acid (LNA)-modi?ed oligonucleotide for hepatitis B virus (HBV) DNA interference in human hepatocytes expressing HBV genomic DNA. In these cells, the LNA-modi?ed oligonucleotides passed ef?ciently across the cell membrane, and lipid-coating facilitated translocation from the cytoplasm to the nucleus. The oligonucleotide specifically targeting HBV DNA clearly interfered with HBV DNA transcription as shown by a block in pregenomic RNA (pgRNA) production. The HBV DNA-targeted oligonucleotide suppressed HBV DNA replication and HBV protein production more ef?ciently than small interfering RNAs directed to the pgRNA. These results demonstrate that fusion with lipid can carry LNA-modi?ed oligonucleotides to the nucleus where they regulate gene expression. Interfering with HBV DNA transcription by LNA-mod-i?ed oligonucleotides has strong potential as a new strategy for HBV inhibition.

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	1. Inhibition of hepatitis B virus (HBV) by LNA-mediated nuclear interference with HBV DNA transcription
	Sun Zhen,Chen Hongyan,Lu Daru
	Biology 04 January 2013
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	Abstract:Silencing target genes with small regulatory RNAs is widely used to investigate gene function and therapeutic drug development. Recently,triplex-based approaches have provided another attractive means to achieve targeted gene regulation and gene manipulation at the molecular and cellular levels. Nuclear entry of oligonucleotides and enhancement of their af?nity to the DNA targets are key points of such approaches. In this study, we developed lipid-based transport of a locked-nucleic-acid (LNA)-modi?ed oligonucleotide for hepatitis B virus (HBV) DNA interference in human hepatocytes expressing HBV genomic DNA. In these cells, the LNA-modi?ed oligonucleotides passed ef?ciently across the cell membrane, and lipid-coating facilitated translocation from the cytoplasm to the nucleus. The oligonucleotide specifically targeting HBV DNA clearly interfered with HBV DNA transcription as shown by a block in pregenomic RNA (pgRNA) production. The HBV DNA-targeted oligonucleotide suppressed HBV DNA replication and HBV protein production more ef?ciently than small interfering RNAs directed to the pgRNA. These results demonstrate that fusion with lipid can carry LNA-modi?ed oligonucleotides to the nucleus where they regulate gene expression. Interfering with HBV DNA transcription by LNA-mod-i?ed oligonucleotides has strong potential as a new strategy for HBV inhibition.
	TO cite this article:Sun Zhen,Chen Hongyan,Lu Daru. Inhibition of hepatitis B virus (HBV) by LNA-mediated nuclear interference with HBV DNA transcription[OL].[ 4 January 2013] http://en.paper.edu.cn/en_releasepaper/content/4511934


	2. Cloning and purifying of Soluble amyloid precursor protein and its modulating MAPK signal pathway
	CHEN Keping,DOU Fei
	Biology 04 January 2013
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	Abstract:It is well known that the proteolytic processing of the amyloid precursor protein (APP) release β-amyloid (Aβ), which plays a central role in the pathogenesis of Alzheimer's disease (AD). Understanding APP processing is important for reducing Aβ levels in AD therapeutic strategy. Soluble amyloid precursor protein (sAPPa) is a proteolyte of APP cleavage by α-secretase. The significance of the cleavage and the physiological functions of sAPPα are poorly understood. In this study, we constructed the stable cell lines expressing sAPPα fusion protein and purified with tandem affinity purification (TAP) technology. We also found sAPPα could modulate the MAPK signal pathway involving in NCAM-induced neurite outgrowth, likes the full length APP.
	TO cite this article:CHEN Keping,DOU Fei. Cloning and purifying of Soluble amyloid precursor protein and its modulating MAPK signal pathway[OL].[ 4 January 2013] http://en.paper.edu.cn/en_releasepaper/content/4509550

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	3. Sexual dimorphism of steroidogenesis regulated by GnIH in the Goldfish, Carassius auratus
	QI Xin,ZHOU Wenyi,LU Danqi,LI Shuisheng,LIU Xiaochun,ZHANG Yong,LIN Haoran
	Biology 22 December 2012
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	Abstract:Gonadotrophin-inhibitory hormone (GnIH) has pronounced inhibitory effects on reproduction in a number of species. GnIH suppresses gonadotropin synthesis/release at the hypothalamic and pituitary levels; however, increasing evidences suggested a putative function on the gonad. In this study, we have demonstrated the localization of receptors for GnIH in ovary and testis of goldfish. In situ hybridization showed that goldfish GnIHRs were localized only in the oocytes before cortical alveolus stage, and were localized in the interstitial tissue of testis. Implantation of goldfish GnIH peptides reduced the serum estradiol level in the female goldfish, but enhanced serum testosterone level in the male goldfish. Besides, injection of goldfish GnIH peptides increased the expression of StAR and 3bHSD mRNA and decreased the expression of CYP19 mRNA significantly in testis, while showed no effect on these genes in ovary. In addition, in vitro results showed that goldfish GnIH peptides not only increased the expression of StAR, 3bHSD and decreased CYP19 mRNA, but also increased the expression of FSHR and LHR mRNA in testis cells, but showed no effect on the expression of detected genes in ovary cells. Thus, we believe that GnIH could contribute to the sexual dimorphism of steroidogenesis in goldfish.
	TO cite this article:QI Xin,ZHOU Wenyi,LU Danqi, et al. Sexual dimorphism of steroidogenesis regulated by GnIH in the Goldfish, Carassius auratus[J].


	4. One new subgenus and one record species of Rhamphomyia (Diptera, Empididae) from China
	YANG Ding,LIU Xingyue
	Biology 30 October 2012
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	Abstract:A new subgenus, Sinorhamphomya subgen. nov., of the genus Rhamphomyia from Palaearctic Region is discribed. One species, Rhamphomyia insignis Loew, is reported from China for the first time. The new subgenus is charaterized by the following features: male eyes contiguous on frons; female eyes widely separated on frons, frons with 6 orb; antennal scape distinctly longer than pedicel, first flagellomere much elongated, style about 1/4 as long as first flagellomere; acr irregularly biseriate, short hair-like; dc irregularly biseriate, short hair-like, 1 long posteriormost dc; prosternum with setulae on lateral parts and lower front parts; dical cell in female much elongated discal cell; anal vein reaching wing margin; male abdominal tergites 6-7 modified, each with one pair of angular lateral processes. It is distributed in the Palaearctic Region with only 1 known species. The new record species is characterized by the legs yellow except extreme tips of mid and hind femora blackish, tip of hind tibia black, and all tarsi entirely black, male cercus rather wide basally and long finger-like apically. It is distributed in China and Russia.
	TO cite this article:YANG Ding,LIU Xingyue. One new subgenus and one record species of Rhamphomyia (Diptera, Empididae) from China[OL].[30 October 2012] http://en.paper.edu.cn/en_releasepaper/content/4492823


	5. PCR and magnetic bead-mediated target capture for the isolation of short interspersed nucleotide elements in fishes
	Liu Dong,Zhu Guoli,Tang Wenqiao,Yang Jinquan,Guo Hongyi
	Biology 06 September 2012
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	Abstract:Short interspersed nucleotide elements (SINEs), a type of retrotransposon, are widely distributed in various genomes with multiple copies arranged in different orientations, and cause changes to genes and genomes during evolutionary history. This can provide the basis for determining genome diversity, genetic variation and molecular phylogeny, etc. SINE DNA is transcribed into RNA by polymerase III from an internal promoter, which is composed of two conserved boxes, box A and box B. Here we present an approach to isolate novel SINEs based on these promoter elements. Box A of a SINE is obtained via PCR with only one primer identical to box B (B-PCR). Box B and its downstream sequence are acquired by PCR with one primer corresponding to box A (A-PCR). The SINE clone produced by A-PCR is selected as a template to label a probe with biotin. The full-length SINEs are isolated from the genomic pool through complex capture using the biotinylated probe bound to magnetic particles. Using this approach, a novel SINE family, Cn-SINE, from the genomes of Coilia nasus, was isolated. The members are 180-360 bp long. Sequence homology suggests that Cn-SINEs evolved from a leucine tRNA gene. This is first report of a tRNALeu-related SINE obtained without the use of a genomic library or inverse PCR. These results provide new insights into the origin of SINEs.
	TO cite this article:Liu Dong,Zhu Guoli,Tang Wenqiao, et al. PCR and magnetic bead-mediated target capture for the isolation of short interspersed nucleotide elements in fishes[OL].[ 6 September 2012] http://en.paper.edu.cn/en_releasepaper/content/4488796


	6. MaHog1, a Hog1-type MAP kinase gene, contributes to stress tolerance and virulence of the entomopathogenic fungus, Metarhizium acridum
	Jin Kai,Ming Yue,Xia Yuxian
	Biology 03 September 2012
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	Abstract:Fungal biocontrol agents have great potential in integrated pest management. However, the poor efficacy and secnsitiveness to various adversities have hampered their wide application. In eukaryotic cells, Hog1 kinase play a critical role in stress responses. In this study, MaHog1 (GenBank accession number EFY85878) encodeing a member of the Hog1/Sty1/p38 MAPK family was identified from M. acridum. Targeted gene disruption was adopted to analyze the role of MaHog1 in virulence and tolerance to adverse factors. Mutant depleted for MaHog1 showed increased sensitivity to high osmotic stress, high temperature and oxidative stress and exhibited remarkable resistance to the cell wall disturbing agents. These results suggested that Hog1 kinase has conserved function in regulating multistress responses among fungi, and that MaHog1 might influence the cell wall biogenesis in M. acridum. Bioassays conducted with topical inoculation and intrahaemocoel injection revealed that MaHog1 is required for both the penetration and the postpenetration development of M. acridum. Disruption of MaHog1 resulted in a signi?cant reduction in virulence, likely due to the combination of a decrease in conidial germination, a reduction in appressorium formation, and a decline in growth rate in insect haemolymph, which might be caused by impairing the fungal tolerance to various stresses during infection.
	TO cite this article:Jin Kai,Ming Yue,Xia Yuxian. MaHog1, a Hog1-type MAP kinase gene, contributes to stress tolerance and virulence of the entomopathogenic fungus, Metarhizium acridum[OL].[ 3 September 2012] http://en.paper.edu.cn/en_releasepaper/content/4488394


	7. The effect of centrally administered apelin-13 on interdigestive gastric motility in conscious rats
	LV Shuangyu,QIN Yaojun,WANG Ningbo,YANG Yanjie,CHEN Qiang
	Biology 02 August 2012
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	Abstract:Apelin, a novel neoropeptide, was identified as the endogenous ligand for APJ receptor. The present study was designed to explore the effect apelin-13 on interdigestive gastric motility in conscious rats. Apelin-13 was synthesized by the solid-phase peptide synthesis method. After recovery from the operation of intracerebroventricular (i.c.v.) cannula implantation, the strain gauge transducers were implanted on the serosal surface of the antrum in Wistar rats. The migrating motor complex (MMC) was instantly recorded by the Power-Lab recording system. The results show that, comparing with control group (5.17±0.93), the frequency of the phase Ⅲ-like activity (c/h) of the 10 nmol apelin-13 group (7.67±1.33) and 30 nmol apelin-13 group (8.58±1.66) were both increased, showing a statistically significant difference (P < 0.01, P < 0.01, respectively). However, frequency of the phase Ⅲ-like activity of 3 nmol apelin-13 group (6.83±0.98) had no significant difference (P > 0.05). Comparing with control group (97.41±16.88), the %motor index (MI) of the 10 nmol apelin-13 group (143.93±26.27) and 30 nmol apelin-13 group (155.09±24.02) were both increased, indicating a statistically obvious difference (P < 0.01, P < 0.01, respectively). However, the %MI of 3 nmol apelin-13 group (120.14±3.00) showed no significant difference (P > 0.05). Our study first indicates that i.c.v. injection of apelin-13 increased the interdigestive antrum MMC in conscious rats, which has an important physiological significance for further study the modulatory effect of apelin-13 on the digestive tract motility.
	TO cite this article:LV Shuangyu,QIN Yaojun,WANG Ningbo, et al. The effect of centrally administered apelin-13 on interdigestive gastric motility in conscious rats[OL].[ 2 August 2012] http://en.paper.edu.cn/en_releasepaper/content/4485939


	8. Effects of Wingless / Wnt1 on the Development of Tribolium castaneum
	ZhuoyuLi,YananPeng,BinLi,ChengjunLi
	Biology 25 June 2012
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	Abstract:Known as highly conserved during evolution, the Wnt signaling pathway plays a vital role in regulating animal embryonic axis, embryonic differentiation, and deciding cell polarity and maintaining adult dynamic equilibrium. Mutations or deregulations of its components can cause the occurrence of carcinoma. We studied the role of Wingless/Wnt1 during larva-adult development of the red flour beetle, Tribolium castaneum, with dsRNA-mediated Wingless(Wg)/Wnt1 gene being knocked down. The treated late larvae metamorphosed into pupae with drastically increased wing interval and decreased wing width (P<0.01). The pursuant pupa-adult eclosion was also severely affected and most of pupae died during this period. The qPCR result showed that the mRNA level of Cadherin-like and Smoothened (Smo) genes were up-regulated greatly, and that of armadillo-2 was slightly higher , after Wingless/Wnt1 gene was knocked down. We drew the conclusion that Wnt-1 signaling pathway is closely related to the proper wing development and adult metamorphosis of Tribolium. In addition, the elevated expression of Cadherin-like and Armadillo-2 may be accountable for the reduced wing width and enlarged wing interval caused by wg gene silencing, because those alterations can either enhance cell adhesion or change cell morphology. Importantly, the up-regulation of smo gene indicates that Hedgehog signaling pathway may be affected by the RNAi of wg and involved in the abnormal or lethal phenotypes observed in our experiment.??
	TO cite this article:ZhuoyuLi,YananPeng,BinLi, et al. Effects of Wingless / Wnt1 on the Development of Tribolium castaneum[OL].[25 June 2012] http://en.paper.edu.cn/en_releasepaper/content/4482937


	9. Cloning and Functional characterization of three Superoxide Dismutases genes from halophyte Salicornia europaea and Thellungiella halophila
	Wu Fan ,Mei Yu,Lu Maolong,Juan Li,Wang Rongfu,Wang Xianlei,Sun Qizhen,Jie Zang,Kai Xu,Hu Ji
	Biology 17 April 2012
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	Abstract:In order to study the halophyte salt-tolerance mechanism, we cloned the manganese (Mn) and Cu/Zn superoxide dismutase (SOD) full-length cDNAs from Salicornia europaea by RACE method for the first time, sequence analysis indicated that the MnSOD gene (GenBank accession number: JQ061158) comprises an open reading frame of 699 bp, encoding 233-amino acid polypeptide with a predicted molecular mass of 25.7 kDa. Correspondingly, the Cu/ZnSOD gene (GenBank accession number: JQ061160) consists of an open reading frame of 684 bp which encodes a protein of 228 amino acids with a predicted molecular mass of 23.3 kDa. The prokaryotic expression vectors pET30-SeMSD, pET30-SeCSD and pET30-ThMSD were constructed, and the target proteins were expressed successfully in BL21 Escherichia coli. Through optimization of the inducing concentration of Isopropyl β-D-Thiogalactopyranoside (IPTG), we tested the salt tolerance of these three superoxide dismutases under 6.5% and 7.5% NaCl, and the results demonstrate that the recombinants BL21 (pET30-SeMSD) and BL21 (pET30-ThMSD) show better tolerance to salinity stress in comparison with the control stain BL21 (pET30), but the recombinant BL21 (pET30-SeCSD) has not displayed increased salt tolerance.
	TO cite this article:Wu Fan ,Mei Yu,Lu Maolong, et al. Cloning and Functional characterization of three Superoxide Dismutases genes from halophyte Salicornia europaea and Thellungiella halophila[OL].[17 April 2012] http://en.paper.edu.cn/en_releasepaper/content/4474413


	10. MaKatG1, a bifunctional catalase-peroxidase gene, contributes to virulence and UV tolerance of Metarhizium acridum
	FAN Anni,PENG Guoxiong,XIA Yuxian
	Biology 16 April 2012
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	Abstract:Entomopathogenic fungi are exposed to reactive oxygen species (ROS) produced either as a secondary products derived from a chemical reaction due to exposure to stress (UV, heat) or by the host. However, the mechanism by which fungal pathogens overcome the toxicity of ROS is unclear. In this study, a bifunctional catalase-peroxidase homolog, KatG1, was identified from Metarhizium acridum, a specific pathogenic fungus against locust. The knockout mutant of MaKatG1 exhibited reduced catalase and peroxidase activity and increased susceptibility to H2O2 and menadione, indicating that MaKatG1 has a bifunctional catalase-peroxidase acctivity and plays a role in the oxidative stress response. The germination and appressorium formation rates of the knockout mutant were decreased on locust wings, but were not affected in vitro. Compared with the wild-type, the virulence of the mutant was signifcantly reduced following topical inoculation, but was not affected following injection. These results demonstrated that the MaKatG1 gene plays a role in the early stages of fungal infection including germination and appressorium formation. Furthermore, UV-B tolerance of the mutant was reduced. This study demonstrated that MaKatG1 contributes to virulence and UV-B tolerance.
	TO cite this article:FAN Anni,PENG Guoxiong,XIA Yuxian. MaKatG1, a bifunctional catalase-peroxidase gene, contributes to virulence and UV tolerance of Metarhizium acridum[OL].[16 April 2012] http://en.paper.edu.cn/en_releasepaper/content/4475570

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