Selectively driving cholinergic fibers optically in the thalamic reticular nucleus promotes sleep.

Cholinergic projections from the basal forebrain and brainstem are thought to play important roles in rapid eye movement (REM) sleep and arousal. Using transgenic mice in which channelrhdopsin-2 is selectively expressed in cholinergic neurons, we show that optical stimulation of cholinergic inputs to the thalamic reticular nucleus (TRN) activates local GABAergic neurons to promote sleep and protect non-rapid eye movement (NREM) sleep. It does not affect REM sleep.

SLXL1, a novel acrosomal protein, interacts with DKKL1 and is involved in fertilization in mice.

Background: Spermatogenesis is a complex cellular developmental process which involves diverse families of genes. The Xlr (X-linked, lymphocyte regulated) family includes multiple members, only a few of which have reported functions in meiosis, post-meiotic maturation, and fertilization of germ cells. Slx-like1 (Slxl1) is a member of the Xlr family, whose expression and function in spermatogenesis need to be elucidated.

The complete genome sequence of Clostridium botulinum F str. 230613, insertion sites, and recombination of BoNT gene clusters.

The genomic DNA of Clostridium botulinum F str. 230613 includes a chromosome (3 993 083 bp, 3502 coding sequences (CDs)) and a plasmid (17 531 bp, 25 CDs). The arrangement of the botulinum neurotoxin serotype F (BoNT/F) gene cluster, a 15-kb (or longer) fragment including the bont gene and other relevant genes, and its different insertion sites in C.

Shiga toxin type 2 (Stx2), a potential agent of bioterrorism, has a short distribution and a long elimination half-life, and induces kidney and thymus lesions in rats.

Shiga toxin type 2, a major virulence factor produced by the Shiga toxin-producing Escherichia coli, is a potential toxin agent of bioterrorism. In this study, iodine-125 (125I) was used as an indicator to describe the in vivo Stx2 biodistribution profile. The rats were injected intravenously (i.

Passive protection of purified yolk immunoglobulin administered against Shiga toxin 1 in mouse models.

Shiga toxins produced by Escherichia coli O157:H7 cause a wide spectrum of enteric diseases, such as lethal hemorrhagic colitis and hemolytic uremic syndrome. In this study, the B subunit protein of Shiga toxin type 1 (Stx1) was produced in the E. coli system, was further purified by Ni-column Affinity Chromatography method, and was then used as an immunogen to immunize laying hens for yolk immunoglobulin (IgY) production.

Fine mapping and interaction analysis of a linear rabies virus neutralizing epitope.

A novel human antibody AR16, targeting the G5 linear epitope of rabies virus glycoprotein (RVG) was shown to have promising antivirus potency. Using AR16, the minimal binding region within G5 was identified as HDFR (residues 261-264), with key residues HDF (residues 261-263) identified by alanine replacement scanning. The key HDF was highly conserved within phylogroup I Lyssaviruses but not those in phylogroup II.

Somatic nucleus reprogramming is significantly improved by m-carboxycinnamic acid bishydroxamide, a histone deacetylase inhibitor.

Somatic cell nuclear transfer (SCNT) has shown tremendous potential for understanding the mechanisms of reprogramming and creating applications in the realms of agriculture, therapeutics, and regenerative medicine, although the efficiency of reprogramming is still low. Somatic nucleus reprogramming is triggered in the short time after transfer into recipient cytoplasm, and therefore, this period is regarded as a key stage for optimizing SCNT. Here we report that CBHA, a histone deacetylase inhibitor, modifies the acetylation status of somatic nuclei and increases the developmental potential of mouse cloned embryos to reach pre- and post-implantation stages.

[Recombinant design and expression of human anti-rabies virus ScdsFv].

To constructed the recombinant human anti-rabies virus ScdsFv, cys sites were introduced into framework region (FR) of VH and VL genes which were amplified from human anti-rabies virus ScFv respectively using genetic point mutation technology. Cloned the ScdsFv gene into expression vector pET22b (+) and transformed into E. coli BL21 (DE3).

[Expression of human anti-rabies Scds-Fv fusion protein in E.coli].

Aim: To express the anti-rabies MBP-ScdsFv fusion protein in E.coli and identify it's activity assay.

Methods: A full length of ScdsFv gene was Cloned into prokaryotic vector pMAL-p2x by recombinant DNA, Then the recombinant plasmid pMAL-ScdsFv was transformed to E.

[Screening of botulinum neurotoxin type A-binding peptides by phage display peptide library].

To seek effective inhibitor against BoNT/A, in this study, BoNT/A-binding peptides were screened from phage display peptide library using synthesized and identified mimicry peptides that contained antigenic epitopes as targets. According to the homology of the amino acid sequences of displayed peptides, most had same motifs respectively. ELISA assay confirmed that identified positive clones respectively against P4 and P5 could specifically bind BoNT/A.

[Preparation of alpha-toxin's protective antigen of clostridium perfringens type A and research for its primary immunological protective function].

Induced by 42 degrees C, the recombinant engineering bacterial pBV/cpa408 was highly expressed. After having been pelleted by 80% (NH4)2 SO4 and dialysised, the expressed protein was isolated and purified by the gel filtration choromatography. Then according to an amount of 1.

[The effects of genomic imprinting on animal development and cloning].

In placental mammals, there is a small group of special genes, which are imprinted so that only one of the parental alleles is actually expressed in target cells. This epigenetic process is named genomic imprinting. Till now, about eighty genes are found to be imprinted in mice and human, and imprinting is conserved in ruminant species as well.

[Construction of human phage-displayed scFv library and selection of the ScFv against rabies virus].

Aim: To construct a human phage displayed single chain Fv antibody (scFv) library and screen specific scFv against rabies virus.

Methods: Using a phage-display technique, IgVH and IgVL genes were cloned from peripheral blood lymphocytes from three donors immunized with the WISTAR PM strain vaccine. Genes encoding scFv fragments were constructed by random linkage of VH gene and VL gene by SOE PCR, and then the constructed scFv genes were cloned into phagemid vector and transfected into E.

[Analysis on mutation of adrenoleukodystrophy gene in exon 1 and exon 5].

Objective: To elucidate the molecular mechanism of X-linked adrenoleukodystrophy(ALD) in Chinese.

Methods: Polymerase chain reaction in exon 1, exon 5 and their flanking sequences and direct DNA sequencing of ALD gene were performed in four patients, their mothers and twenty normal individuals as controls.

Results: A splice mutation was identified in the interface of exon 5 and intron 5 (1875 G-->A).