Changchun, China
Changchun, China

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Wei J.,Changchun Normal University | Wei J.,Jilin Agricultural University | Guo N.,Institute of Zoonosis | Guo N.,Jilin University | And 6 more authors.
Polish Journal of Microbiology | Year: 2013

Tuberculosis (TB), affecting one-third of the global population, kills an estimated two to three million people every year. The development of drug resistance is becoming a serious threat to any attempt to control this disease, which underscores the need for new agents targeting Mycobacterium tuberculosis (M. tuberculosis). Osthole (7-methoxy-8-isopentenoxycoumarin) is a coumarin derivative present in many medicinal plants. Previous studies have shown that osthole possesses antimycobacterial effects, however, the action mechanism of osthole is unclear. In the study, we used a commercial oligonucleotide microarray to determine the overall transcriptional response of M. tuberculosis H37Rv triggered by exposure to osthole. Analysis of the microarray data revealed that a total of 478 genes were differentially regulated by osthole. Of these, 241 genes were upregulated, and 237 genes were downregulated. Some of the important genes that were significantly regulated are related to different pathways such as fumarate reductase, class I peroxidase, cell wall, nitrate respiration, and protein synthesis. Real-time quantitative RT-PCR was performed for chosen genes to validate the microarray results. To our knowledge, this genome-wide transcriptomics approach has produced the first insights into the response of M. tuberculosis when exposed to osthole.


Wei J.,Changchun Normal University | Wei J.,Jilin University | Liang J.,Institute of Zoonosis | Shi Q.,Institute of Zoonosis | And 5 more authors.
Brazilian Journal of Microbiology | Year: 2014

Mycobacterium tuberculosis (M. tuberculosis), the causative agent of tuberculosis, still causes higher mortality than any other bacterial pathogen until now. With the emergence and spread of multidrug-resistant (MDR) and extensively drug-resistant (XDR-TB) strains, it becomes more important to search for alternative targets to develop new antimycobacterial drugs. Lupulone is a compound extracted from Hops (Hurnulus lupulus), which exhibits a good antimicrobial activity against M. tuberculosis with minimal inhibitory concentration (MIC) value of 10 μg/mL, but the response mechanisms of lupulone against M. tuberculosis are still poorly understood. In this study, we used a commercial oligonucleotide microarray to determine the overall transcriptional response of M. tuberculosis H37Rv triggered by exposure to MIC of lupulone. A total of 540 genes were found to be differentially regulated by lupulone. Of these, 254 genes were upregulated, and 286 genes were downregulated. A number of important genes were significantly regulated which are involved in various pathways, such as surface-exposed lipids, cytochrome P450 enzymes, PE/PPE multigene families, ABC transporters, and protein synthesis. Real-time quantitative RT-PCR was performed for choosed genes to verified the microarray results. To our knowledge, this genome-wide transcriptomics approach has produced the first insights into the response of M. tuberculosis to a lupulone challenge. © 2014, Sociedade Brasileira de Microbiologia.


Ma L.,Institute of Zoonosis | Ma L.,Jilin University | Liu B.,Jilin University | Jiang Z.,Jilin University | And 2 more authors.
Clinical Rheumatology | Year: 2014

This study is aimed at determining the numbers of circulating Treg and Breg cells in patients with new-onset rheumatoid arthritis and during subsequent drug therapies. Patients were treated orally with 10 mg methotrexate weekly, and 20 mg leflunomide and 60 mg common threewingnut root daily (Lei Gong Teng) for 12 weeks, but received no steroid therapy. Basal measurements were performed of serum C-reactive protein, anticyclic citrullinated peptide antibody, and erythrocyte sedimentation rate, and the numbers of cluster of differentiation CD4+CD25+Foxp3+ T cells, interleukin 10 (IL10)-expressing on CD5+CD1d+ and TIM1+ B cells. Compared with the healthy controls, patients exhibited significantly less numbers of circulating CD19+TIM1+IL10+, CD19+CD5+CD1d+IL10+ B cells and CD4+CD25+Foxp3+ T cells (P < 0.001, all). Drug therapy modulated the balance of different subsets of Breg and Treg cells. The numbers of CD19+TIM1+IL10+ and CD19 +CD5+CD1d+IL10+ B cells correlated positively with the numbers of CD4+CD25+Foxp3+ T cells in these patients (r = 0.707, P = 0.001; r = 0.481, P = 0.007, respectively). The values of DAS28 were negatively correlated with the numbers of CD19+TIM1+IL10+ and CD19+CD5 +CD1d+IL10+ B cells, and CD4 +CD25+Foxp3+ T cells (r = -0.533, P = 0.023; r = -0.442, P = 0.016; and r = -0.444, P = 0.014, respectively). Of note, TIM1+ B cells identified more circulating IL10+ B cells than CD5+CD1d+ B cells. Our data indicate that Breg and Treg cells have a potentially crucial role in controlling disease activity in rheumatoid arthritis patients, and TIM1+ Breg cells may be a viable therapeutic target for these patients. © 2013 Clinical Rheumatology.


Chen F.-Y.,Xiamen University | Liu H.-P.,Xiamen University | Bo J.,Xiamen University | Ren H.-L.,Xiamen University | And 2 more authors.
Fish and Shellfish Immunology | Year: 2010

Although the crab Scylla paramamosain has been cultured in China for a long time, little knowledge is available on how crabs respond to infection by bacteria. A forward suppression subtractive hybridization (SSH) cDNA library was constructed from their hemocytes and the up-regulated genes were identified in order to isolate differentially expressed genes in S. paramamosain in response to bacterial lipopolysaccharide (LPS). A total of 721 clones on the middle scale in the SSH library were sequenced. Among these genes, 271 potentially functional genes were recognized based on the BLAST searches in NCBI and were categorized into seven groups in association with different biological processes using AmiGO against the Gene Ontology database. Of the 271 genes, 269 translatable DNA sequences were predicted to be proteins, and the putative amino acid sequences were searched for conserved domains and proteins using the CD-Search service and BLASTp. Among 271 genes, 179 (66.1%) were annotated to be involved in different biological processes, while 92 genes (33.9%) were classified as an unknown-function gene group. It was noted that only 18 of the 271 genes (6.6%) had previously been reported in other crustaceans and most of the screened genes showed less similarity to known sequences based on BLASTn results, suggesting that 253 genes were found for the first time in S. paramamosain. Furthermore, two up-regulated genes screened from the SSH library were selected for full-length cDNA sequence cloning and in vivo expression study, including Sp-superoxide dismutase (Sp-Cu-ZnSOD) gene and Sp-serpin gene. The differential expression pattern of the two genes during the time course of LPS challenge was analyzed using real-time PCR. We found that both genes were significantly expressed in LPS-challenged crabs in comparison with control. Taken together, the study primarily provides the data of the up-regulated genes associated with different biological processes in S. paramamosain in response to LPS, by which the interesting genes or proteins potentially involved in the innate immune defense of S. paramamosain will be investigated in future. © 2009 Elsevier Ltd. All rights reserved.


Huang W.-B.,Xiamen University | Ren H.-L.,Xiamen University | Ren H.-L.,Institute of Zoonosis | Gopalakrishnan S.,Xiamen University | And 3 more authors.
Fish and Shellfish Immunology | Year: 2010

Mammal caspases have been demonstrated to possess important functions in apoptosis and immune signaling, but there is less knowledge available on abalone caspases. In the present study, a molluscan caspase gene, abCaspase, was cloned for the first time from the variously colored abalone (Haliotis diversicolor) and its full-length cDNA sequence was 2427 bp, with a 1008 bp of open reading frame encoding a protein of 336 aa. The molecular mass of the deduced protein was approximately 36.97 kDa with an estimated pI of 5.28. The predicted amino acid sequence of abCaspase contained two domains of p20 and p10 which were conserved in the caspase family, including the cysteine active site pentapeptide "QSCRG" and the histidine active site signature "HTVYDCVVVIFLTHG". Homology analysis showed that abCaspase shared high similarity with apoptotic caspases and it was grouped together with vertebrate caspase-8s and caspase-10s using phylogenetic analysis, suggesting that abCaspase belonged to a typical apoptotic caspase and might possess the characteristic of human caspase-8 and -10. The mRNA transcripts of abCaspase were widely distributed in various tissues of H. diversicolor. Expression of the abCaspase gene was significantly induced in the tissues tested, especially in the hemocytes, gill and mantle with bacterial challenge. This study suggested that abCaspase may be an initiator caspase associated with the induction of apoptosis which is potentially involved in the immune defense of H. diversicolor. © 2009 Elsevier Ltd. All rights reserved.


Chu X.,Institute of Zoonosis | Ci X.,Institute of Zoonosis | Wei M.,Institute of Zoonosis | Yang X.,Institute of Zoonosis | And 6 more authors.
Journal of Agricultural and Food Chemistry | Year: 2012

Licochalcone A (Lico A), a flavonoid found in licorice root (Glycyrrhiza glabra), is known for its antimicrobial activity and its reported ability to inhibit cancer cell proliferation. In the present study, we found that Lico A exerted potent anti-inflammatory effects in in vitro and in vivo models induced by lipopolysaccharide (LPS). The concentrations of TNF-α, interleukin (IL)-6, and IL-1β in the culture supernatants of RAW 264.7 cells were determined at different time points following LPS administration. LPS (0.5 mg/kg) was instilled intranasally (i.n.) in phosphate-buffered saline to induce acute lung injury, and 24 h after LPS was given, bronchoalveolar lavage fluid was obtained to measure pro-inflammatory mediator and total cell counts. The phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear factor-κB (NF-κB) p65 protein was analyzed by Western blotting. Our results showed that Lico A significantly reduced the amount of inflammatory cells, the lung wet-to-dry weight (W/D) ratio, protein leakage, and myeloperoxidase activity and enhances oxidase dimutase activity in mice with LPS-induced acute lung injury (ALI). Enzyme-linked immunosorbent assay results indicated that Lico A can significantly down-regulate TNF-α, IL-6, and IL-1β levels in vitro and in vivo, and treatment with Lico A significantly attenuated alveolar wall thickening, alveolar hemorrhage, interstitial edema, and inflammatory cells infiltration in mice with ALI. In addition, we further demonstrated that Lico A exerts an anti-inflammation effect in an in vivo model of acute lung injury through suppression of NF-κB activation and p38/ERK MAPK signaling in a dose-dependent manner. © 2012 American Chemical Society.


Zhou Y.,Institute of Zoonosis | Li Y.,Institute of Zoonosis | Lu S.,Institute of Zoonosis | Ren H.,Institute of Zoonosis | And 6 more authors.
Sensors and Actuators, B: Chemical | Year: 2010

A competitive format immunoassay based on a gold nanoparticle-labelled monoclonal antibody (Mab) for the rapid detection of tetrodotoxin (TTX) in puffer fish tissues was developed. In the assay, two antibodies were employed. The first antibody supplied in sample pad was as detector reagent and the secondary antibody immobilized on a diagnostic cellulose nitrate membrane was as capture reagent. TTX in tissue sample competed with immobilized TTX-BSA to bind with gold nanoparticle-labelled Mab. The mobile complex (gold nanoparticle-Mab-TTX) can be captured by the secondary antibody but cannot be captured by TTX-BSA (test line). The color density of the test line correlated with the concentration of TTX in sample in the range 40-8000 ng mL-1. The visual detection limit of TTX-spiked samples was found to be 40 ng mL-1. The assay time of the qualitative test based on the visual evaluation of results for TTX detection was less than 10 min, and the assay performance did not require any equipment. This method can be applied as a screening method for rapid testing of TTX on site. © 2010 Elsevier B.V. All rights reserved.


Zhou Y.,Institute of Zoonosis | Li Y.-S.,Institute of Zoonosis | Pan F.-G.,Institute of Zoonosis | Zhang Y.-Y.,Institute of Zoonosis | And 5 more authors.
Food Chemistry | Year: 2010

Brevetoxin B (PbTx-2) was covalently linked to carrier protein bovine serum albumin and human gamma globulin. A monoclonal antibody against PbTx-2, which showed high cross-reactivity values with PbTx-1, PbTx-3 and PbTx-9 (more than 89%) was obtained from ascites and some characteristics of monoclonal antibody were studied. An direct competitive enzyme-linked immunosorbent assay (ELISA) for detection of PbTxs was developed, which showed an IC50 value of 5.3 ng mL-1 with a detection limit of 0.6 ng well-1. The recoveries of PbTxs from cockle (88.4%-102.3%) and oyster (89.4%-104.3%) demonstrated that the matrices of cockle and oyster where PbTxs are found do not interfere with the assay. The newly developed competitive ELISA appears to be a reliable and useful method for mass monitoring of PbTxs in mollusk. © 2009 Elsevier Ltd. All rights reserved.

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