Author Correction: Linc-DYNC2H1-4 promotes EMT and CSC phenotypes by acting as a sponge of miR-145 in pancreatic cancer cells.

After publication of this article, the below errors were noticed:1. The SOX2 primer is incorrect in Table S2.2. The Poly(T) adaptor sequence of reverse transcription for miR-145 detection is missing in Table S2.This error did not impact the conclusions of the article. We apologize for any confusion or inconvenience to the readers.An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Lipidomics characterization of the alterations of Trichoderma brevicompactum membrane glycerophospholipids during the fermentation phase.

The biological membrane lipid composition has been demonstrated to greatly influence the secretion of secondary metabolites. This study was conducted to investigate the periodical alterations of whole cellular lipids and their associations with secondary products in Trichoderma brevicompactum. An electrospray ionization-mass spectrometry-based lipidomics strategy was used to acquire the metabolic profiles of membrane lipids during fermentation. Univariate analyses showed that most fungi glycerophospholipids were significantly altered at the early phase compared with the late phase. In addition, correlation analyses showed high correlations between phosphatidylcholine alterations and fermentation duration. In addition, the fermentation-associated alterations of phosphatidylcholines were found to be in accordance with the degrees of unsaturation of acyl-chains. Harzianum A reached a maximum on the 12th day, while trichodermin and 6-pentyl-2H-pyran-2-one showed the highest abundances on the 9th day, both of which were inclined to correlate with the alterations of phosphatidylcholines and phosphatidylethanolamines, respectively. These findings demonstrated that the alterations of the membrane lipid species in Trichoderma spp. were associated with the fermentation phases and might influence the secretion of specific secondary products, which may be useful in studying the optimization of secondary products in Trichoderma spp.

Linc-DYNC2H1-4 promotes EMT and CSC phenotypes by acting as a sponge of miR-145 in pancreatic cancer cells.

The acquisition of epithelial-mesenchymal transition (EMT) and/or existence of a sub-population of cancer stem-like cells (CSC) are associated with malignant behavior and chemoresistance. To identify which factor could promote EMT and CSC formation and uncover the mechanistic role of such factor is important for novel and targeted therapies. In the present study, we found that the long intergenic non-coding RNA linc-DYNC2H1-4 was upregulated in pancreatic cancer cell line BxPC-3-Gem with acquired gemcitabine resistance. Knockdown of linc-DYNC2H1-4 decreased the invasive behavior of BxPC-3-Gem cells while ectopic expression of linc-DYNC2H1-4 promoted the acquisition of EMT and stemness of the parental sensitive cells. Linc-DYNC2H1-4 upregulated ZEB1, the EMT key player, which led to upregulation and downregulation of its targets vimentin and E-cadherin respectively, as well as enhanced the expressions of CSC makers Lin28, Nanog, Sox2 and Oct4. Linc-DYNC2H1-4 is mainly located in the cytosol. Mechanically, it could sponge miR-145 that targets ZEB1, Lin28, Nanog, Sox2, Oct4 to restore these EMT and CSC-associated genes expressions. We proved that MMP3, the nearby gene of linc-DYNC2H1-4 in the sense strand, was also a target of miR-145. Downregulation of MMP3 by miR-145 was reverted by linc-DYNC2H1-4, indicating that competing with miR-145 is one of the mechanisms for linc-DYNC2H1-4 to regulate MMP3. In summary, our results explore the important role of linc-DYNC2H1-4 in the acquisition of EMT and CSC, and the impact it has on gemcitabine resistance in pancreatic cancer cells.

Studies on the pathogenesis of a Chinese strain of bovine parainfluenza virus type 3 infection in Balb/c mice.

To date, three genotypes A, B, and C of bovine parainfluenza virus type 3 (BPIV3) have been isolated from cattle and only limited studies on the pathogenesis of the genotype A of BPIV3 infection in calves and laboratory animals have been conducted. The pathogenesis of the genotypes B and C of BPIV3 infection in calves and laboratory animals have not been reported. To alleviate the difficulties associated with sourcing suitable calves for infection studies, the establishment of BPIV3 infection model using laboratory model animals could aid in increasing the knowledge of the pathogenesis of this virus. Therefore thirty Balb/c mice were intranasally inoculated with a Chinese BPIV3 strain SD0835 which was classified as genotype C. Virus replications in mice were demonstrated by using virus isolation and titration, immunofluorescent staining, and immunohistochemistry and had occurred in the respiratory tissues as early as 24h after intranasal inoculation. The results of immunofluorescent staining and IHC implicated that the lungs and tracheas might be the major tissues in which the SD0835 infected and replicated. The histopathologic examinations revealed that alveoli septa thickening and focal cellulose pneumonia were seen in the lungs of experimentally infected mice. The aforementioned results indicated that the SD0835 of the genotype C was pathogenic to Balb/c mice and the mouse infection model could cast light on the genotype C of BPIV3 infection process and pathogenesis.

Isolation, identification, and complete genome sequence of a bovine adenovirus type 3 from cattle in China.

BACKGROUND: Bovine adenovirus type 3 (BAV-3) belongs to the Mastadenovirus genus of the family Adenoviridae and is involved in respiratory and enteric infections of calves. The isolation of BAV-3 has not been reported prior to this study in China. In 2009, there were many cases in cattle showing similar clinical signs to BAV-3 infection and a virus strain, showing cytopathic effect in Madin-Darby bovine kidney cells, was isolated from a bovine nasal swab collected from feedlot cattle in Heilongjiang Province, China. The isolate was confirmed as a bovine adenovirus type 3 by PCR and immunofluorescence assay, and named as HLJ0955. So far only the complete genome sequence of prototype of BAV-3 WBR-1 strain has been reported. In order to further characterize the Chinese isolate HLJ0955, the complete genome sequence of HLJ0955 was determined. RESULTS: The size of the genome of the Chinese isolate HLJ0955 is 34,132 nucleotides in length with a G+C content of 53.6%. The coding sequences for gene regions of HLJ0955 isolate were similar to the prototype of BAV-3 WBR-1 strain, with 80.0-98.6% nucleotide and 87.5-98.8% amino acid identities. The genome of HLJ0955 strain contains 16 regions and four deletions in inverted terminal repeats, E1B region and E4 region, respectively. The complete genome and DNA binding protein gene based phylogenetic analysis with other adenoviruses were performed and the results showed that HLJ0955 isolate belonged to BAV-3 and clustered within the Mastadenovirus genus of the family Adenoviridae. CONCLUSIONS: This is the first study to report the isolation and molecular characterization of BAV-3 from cattle in China. The phylogenetic analysis performed in this study supported the use of the DNA binding protein gene of adenovirus as an appropriate subgenomic target for the classification of different genuses of the family Adenoviridae on the molecular basis. Meanwhile, a large-scale pathogen and serological epidemiological investigations for BVA-3 infection might be carried out in cattle in China. This report will be a good beginning for further studies on BAV-3 in China.

Isolation and genetic characterization of bovine parainfluenza virus type 3 from cattle in China.

Bovine parainfluenza virus type 3 (BPIV3) is one of the most important of the known viral respiratory pathogens of both young and adult cattle. However BPIV3 has not been detected or isolated in China prior to this study. In 2008, four BPIV3 strains were isolated with MDBK cells from cattle in China and characterized by RT-PCR, nucleotide sequence analysis, transmission electron microscope observation, hemadsorption and hemagglutination tests. Nucleotide phylogenetic analysis of partial hemagglutinin-neuraminidase (HN) gene for four isolates and the complete genome for the SD0835 isolate implicated that the four Chinese BPIV3 strains were distinct from the previously reported genotype A (BPIV3a) and genotype B (BPIV3b) and might be a potentially new genotype, which was tentatively classified as genotype C (BPIV3c). This is the first study to report the isolation and genetic characterization of BPIV3 from cattle in China.

Genotyping of bovine viral diarrhea viruses from cattle in China between 2005 and 2008.

Eighteen bovine viral diarrhea viruses (BVDV) from cattle in China between 2005 and 2008 were genetically typed by sequencing of the 5'-untranslated region (5'-UTR) of the viral genome and for selected isolates the N(pro) region. Phylogenetic reconstructions indicated that all of the 18 BVDV positive samples examined in this work clustered within the BVDV type 1 genotype. Of the 15 previously described subgenotypes of BVDV1 (1a-1o), 12 of the samples examined in this work clustered with the Chinese BVDV ZM-95 strain of pig origin, which was the prototype of BVDV1m, while 2 samples clustered with the BVDV1b. But 4 samples formed a separate group appearing to be a potentially new subgenotype, which was tentatively typed as "BVDV1p". Based on these results there appears to be highly genetic variation within the Chinese BVDV field isolates. As well, the phylogenetic reconstructions indicate that the clustering of the Chinese BVDV1m subgenotype in the phylogenetic tree is a result of geographic isolation. The information obtained from this work will be useful when carrying out epidemiological surveys of BVDV detected in China, especially for the BVDV1m detection in Chinese cattle.

[Construction and identification of recombinant BHV-1 expressing foot and mouth disease virus VP1 gene].

OBJECTIVE: In order to construct the recombinant bovine hepervirus-1 (BHV-1) which expressed foot and mouth disease virus (FMDV) VP1 gene, we constructed a BHV-1 gE gene transfer vector by inserting the synthetic VP1 gene of FMDV (O/China/99) under the immediate-early promoter of cytomegalovirus. METHODS: The mixtures of parental virus (BHV-1/gE(-)/LacZ+) DNA and transfer vector was transfected into bovine turbinate cells using calcium phosphate-mediated transfection. Then the propagated viruses were harvested. The recombinant BHV-1 (designated BHV-1/gE(-)/VP1) was obtained by selection for white virus plaques. RESULTS: PCR results showed that VP1 gene was successfully inserted into the genome of BHV-1/gE(-). The expression of VP1 in infected cells was proved by indirect immunofluorescence assay and Western blotting. CONCLUSION: The research provided a basis for development of BHV-1 vector vaccines for FMD and other important bovine infectious diseases.

Construction of a recombinant BHV-1 expressing the VP1 gene of foot and mouth disease virus and its immunogenicity in a rabbit model.

Foot-and-mouth disease (FMD) and infectious bovine rhinotracheitis (IBR) are two important infectious diseases of cattle. Using bovine herpesvirus type 1 (BHV-1) as a gene delivery vector for development of live-viral vaccines has gained widespread interest. In this study, a recombinant BHV-1 was constructed by inserting the synthetic FMDV (O/China/99) VP1 gene in the the gE locus of BHV-1 genome under the control of immediately early gene promoter of human cytomegalovirus (phIE CMV) and bovine growth hormone polyadenylation (BGH polyA) signal. After homologous recombination and plaque purification, a recombinant virus named BHV-1/gE(-)/VP1 was acquired and identified. The immunogenicity was confirmed in a rabbit model by virus neutralization test and enzyme-linked immunosorbent assay (ELISA). The result indicated that the BHV-1/gE(-)/VP1 has the potential for being developed as a bivalent vaccine for FMD and IBR.