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Open access

Guanying Wang, Xinran Li, Renli Jiang, Yue Li, Xiaojing Fan, Yu Zheng and Li Gao

Abstract

The purpose of the study was to define transient changes in the concentration of inflammatory biomarkers and cartilage biomarkers in the synovial fluid of joints following experimentally induced acute equine synovitis. Acute synovitis was induced in eight skeletally mature mares by a sterile intra-articular injection of 1 mL of phosphate-buffered saline (PBS) containing 0.5 ng of lipopolysaccharide (LPS). The solution was injected into the right middle carpal joint. One mL of sterile PBS was injected into the left control joint. Synovial fluid was obtained at the baseline level and at 8, 24, and 168 h after injection. The levels of inflammatory biomarkers-prostaglandin E2 (PGE2), interleukin 1β (IL-1β), and tumour necrosis factor-α (TNF-α), and cartilage turnover biomarkers-collagenase-cleavage neoepitope of type II collagen (C2C) and C-terminal crosslinked telopeptide type II collagen (CTX-II) were detected with proper assays. Single injections of LPS raised the number of synovial white blood cells and concentrations of total protein, PGE2, IL-1β, TNF-α, C2C, and CTX-II. PGE2 and IL-1β rose sharply at 8 h, while TNF-α increased steadily through 8 h and 24 h, at that point; these three factors returned to the baseline level by 168 h. The time course of C2C and CTX-II concentrations peaked sharply at 24 h, and continued to be significantly elevated over the baseline level even at 168 h. Injections of LPS into the joints led to a temporal inflammatory response, which in turn increased local release of inflammatory biomarkers and significantly altered the concentrations of cartilage markers in the synovial fluid.

Open access

Li-Wei Gao and Guo-Liang Wang

Abstract

Lung cancer (LC), which includes small-cell lung carcinoma (SCLC) and non-small-cell lung carcinoma (NSCLC), is common and has a high fatality rate. This study aimed to reveal the prognostic mechanisms of LC. GSE30219 was extracted from the Gene Expression Omnibus (GEO) database, and included 293 LC samples and 14 normal lung samples. Differentially expressed genes (DEGs) were identified using the Limma package, and subjected to pathway enrichment analysis using DAVID. MicroRNAs (miRNAs) targeting the DEGs were predicted using Webgestalt. Cytoscape software was used to build a protein-protein interaction (PPI) network and to identify significant network modules. Survival analysis was conducted using Survminer and Survival packages, and validation was performed using The Cancer Genome Atlas (TCGA) dataset. The good and poor prognosis groups contained 518 DEGs. miR-190, miR-493, and miR-218 for the upregulated genes and miR-302, miR-200, and miR-26 for the downregulated genes were predicted. Three network modules (module 1, 2, and 3) were identified from the PPI network. CDK1, MCM10, and NDC80 were the core nodes of module 1, 2, and 3, respectively. In module 1, CDK1 interacted with both CCNB1 and CCNB2. Additionally, CDK1, CCNB1, CCNB2, MCM10, and NDC80 expression levels correlated with clinical survival and were identified as DEGs in both GSE30219 and the TCGA dataset. miR-190, miR-493, miR-218, miR-200, and miR-302 might act in LC by targeting the DEGs. CDK1, CCNB1, CCNB2, MCM10, and NDC80 might also influence the prognosis of LC.

Open access

Renli Jiang, Li Gao, Guanying Wang, Xinran Li, Yue Li, Xiaojing Fan, Xu Liu, Jinglu Wang, Yu Zhang, Xiangxing Kong and Jianhua Xiao

Abstract

Horses (n = 20) were divided into 2 groups: oligofructose (OF)-induced equine laminitis group (group OF; n = 11) which received 10 g/kg b.w. of OF dissolved in 4 L water via nasogastric intubation, and control group (NS; n = 9) which received 4 L of saline. Blood was collected at 4 h intervals over 72 h study period and analysed by ELISA, kinetic limulus amoebocyte lysate assay, and glucose-oxidase methods. The level of insulin changed significantly in horses which received OF (P < 0.01); there was a significant negative correlation between the level of adiponectin and insulin over time. The results suggested that insulin may play an important role in the development of OF-induced equine laminitis by altering the level of endothelin-1 and nitric oxide.

Open access

Xinran Li, Renli Jiang, Guanying Wang, Yue Li, Xiaojing Fan, Xu Liu, Jinglu Wang, Jialiang Pan and Li Gao

Abstract

The study was conducted on 24 Mongolian horses, with oligofructose-induced equine laminitis (10 g/kg b.w.). The objective of the study was to investigate the relationships among matrix metalloproteinase 2 (MMP-2), P38 mitogen-activated protein kinases (P38 MAPK), tissue inhibitor of metalloproteinase 2 (TIMP-2), lipopolysaccharides (LPS), and tumour necrosis factor-α (TNF-α) during acute developmental phase of laminitis, and to determine whether there are any characteristic tendencies. Moreover, plasma concentrations of LPS and TNF-α were measured in order to determine the time of leukocytes’ activation. Eleven of the 12 horses showed clinical signs of laminitis. The contents of MMP-2 and P38 MAPK increased significantly from 8 h to 64 h, and the content of TIMP-2 decreased significantly at the same time. Plasma LPS concentrations increased significantly between 8 h and 20 h and reached a peak of 0.024 ± 0.009 EU/mL (equivalent to 3.04 ± 1.19 pg/mL) at 12 h. TNF-α concentration increased between 20 h and 36 h. This data indicates that MMP-2 plays an important role during the early acute developmental phase of oligofructose-induced equine laminitis.

Open access

Tian-wen Ma, Yue Li, Guan-ying Wang, Xin-ran Li, Ren-li Jiang, Xiao-peng Song, Zhi-heng Zhang, Hui Bai, Xin Li and Li Gao

Abstract

Introduction: The study aimed to clarify the changes in the concentration of inflammatory mediators, proteases, and cartilage degradation biomarkers in the synovial fluid of joints in an equine osteoarthritis model.

Material and Methods: Osteoarthritis was induced in eight Mongolian horses by a sterile intra-articular injection of amphotericin B, which was injected into the left carpal joint in a dose of 2 mL (25 mg/mL). The control group comprised five horses which were injected with an equal dose of sterile physiological saline into the left carpal joint. Synovial fluid was obtained at baseline and every week after injection. Test methods were based on ELISA.

Results: In the course of the osteoarthritis, the concentration of biomarkers in joint synovial fluid showed an increasing trend. IL-1, IL-6, MMP-9, MMP-13, ADAMTS-5, CS846, GAG, HA, CTX-II, and COMP concentrations sharply increased before the onset of significant symptoms of lameness, whereas TNF-α, MMP-2, and MMP-3 concentrations rose sharply after the occurrence of such symptoms.

Conclusion: The results obtained confirm that the concentrations of IL-1, IL-6, MMP-9, MMP-13, ADAMTS-5, CS846, GAG, HA, CTX-II and COMP increase substantially in equine osteoarthritis, which provides a theoretical basis for the rapid diagnosis of the disease.

Open access

Yi-Ming Zhang, Dong-Xu Yu, Bai-Shuang Yin, Xin-Ran Li, Li-Na Li, Ya-Nan Li, Yu-Xin Wang, Yu Chen, Wen-Han Liu and Li Gao

Abstract

Introduction: Xylazine, a type of α2-adrenoceptors, is a commonly used drug in veterinary medicine. Xylazine-induced changes in the content of amino acid neurotransmitters – glycine (Gly) and aspartic acid (Asp), in different brain regions and neurons were studied.

Material and Methods: Wistar rats were administered 50 mg/kg or 70 mg/kg of xylazine by intraperitoneal injection. In addition, in vitro experiments were conducted, in which neurons were treated with 15 μg/mL, 25 μg/mL, 35 μg/mL, and 45 μg/mL of xylazine. Test methods were based on the enzyme-linked immunosorbent assays (ELISA).

Results: During anaesthesia, Asp levels in each brain area were significantly lower compared to the control group. Except for the cerebrum, levels of Gly in other brain areas were significantly increased during the anaesthesia period. In vitro, xylazine-related neuron secretion of Gly increased significantly compared to the control group at 60 min and 90 min. Moreover, xylazine caused a significant decrease in the levels of Asp secreted by neurons at 20 min, but gradually returned to the level of the control group.

Conclusion: The data showed that during anaesthesia the overall levels of Asp decreased and overall levels of Gly increased. In addition, the inhibitory effect of xylazine on Asp and the promotion of Gly were dose-dependent. Our data showed that different effects of xylazine on excitatory and inhibitory neurotransmitters provided a theoretical basis for the mechanism of xylazine activity in clinical anaesthesia.

Open access

Zhigang Zhang, Jianguo Wang, Ruifeng Gao, Weiqian Zhang, Xinwei Li, Guowen Liu, Xiaobing Li, Zhe Wang and Xinglin Zhu

Abstract

The objective of the study was to determine expression of gene of insulin receptor (INSR) in adipose tissue of postpartum dairy cows fed diets containing different amounts of energy at the antepartum period. Healthy pregnant dairy cows (n=45) on 21st d of the antepartum were divided into three groups differing in diet composition, namely: control group fed a normal diet, high energy group fed a high energy diet, and low energy group fed a low energy diet. Twenty-one days after parturition, INSR gene expression in adipose tissue was determined by internally controlled reverse transcriptase PCR. The level of INSR mRNA in adipose tissues of cows fed the high energy diet was substantially lower than that in cows fed normal or low energy diets. A relatively higher level of INSR mRNA in the adipose tissue of cows fed low energy diet may be beneficial for gluconeogenesis and lipogenesis, which can relieve an energy negative balance. Reduced level of INSR mRNA in adipose tissue of cows fed high energy diet indicates that the response to insulin has significantly decreased.

Open access

Yong Zhang, S. G. Zhang, L.W. Qi, B. Liu, J. M. Gao, C. B. Chen, X. L. Li and Wenqin Song

Abstract

The chromosome microdissection, cloning and painting technology has evolved into an efficient tool for genomic research. Application of these techniques has rarely been applied for forest plants, largely due to the difficulty of chromosome preparation. The present study was performed to establish a method for single chromosome microdissection, cloning and painting in forest plants using poplar (Populus tremula) as a model. An individual chromosome 1 was microdissected from the metaphase spreads of poplar root-tip cells with fine glass needle controlled by a micromanipulator. The dissected chromosome was amplified in vitro by the Sau3A linker adaptor mediated PCR (LA-PCR) technique, by which 200bp to 3,000bp smear DNA fragments were obtained. Then, the second round PCR products from the single chromosome 1 were cloned into T-easy vectors to generate a DNA library of the chromosome 1. Approximately 3 x 105 recombinant clones were obtained. The second round PCR products were used as a complex probe mixture for fluorescent in situ hybridization (FISH) on the metaphase spreads of poplar. Hybridization signals were observed, mainly, along the entire chromosome 1, at the same time, signals were also present on telomeric and centromeric regions of other chromosomes. Therefore, this research suggests that chromosome microdissection, cloning and painting of the single small chromosome in forest plants are feasible.