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1.
Int J Mol Sci ; 16(3): 4997-5013, 2015 Mar 04.
Article in English | MEDLINE | ID: mdl-25749476

ABSTRACT

MicroRNAs are small non-coding RNA molecules that are important regulators of gene expression at the post-transcriptional level. miRNAs impact the processes of cell proliferation, differentiation and apoptosis. Thus, the regulation of miRNA expression profiles associated with mastitis will be conducive for its control. In this study, Staphylococcus aureus (S. aureus) was administered to the mammary gland of Chinese Holstein cows to construct a bacteria-type mastitis model. Total RNA was isolated from bovine mammary gland tissue samples from the S. aureus-induced mastitis group and controls. miRNAs were analyzed using Solexa sequencing and bioinformatics processing for the experimental group and control group. Two miRNA libraries were constructed respectively. A total of 370 known bovine miRNAs and 341 novel mi RNAs were detected for the S. aureus and 358 known bovine miRNAs and 232 novel miRNAs for control groups. A total of 77 miRNAs in the S. aureus group showed significant differences compared to the control group. GO (Gene Ontology) analysis showed these target genes were involved in the regulation of cells, binding, etc., while KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis showed that these genes were enriched in endocytosis, and olfactory transduction pathways involved in cancer. These results provide an experimental basis to reveal the cause and regulatory mechanism of mastitis and also suggest the potential of miRNAs to serve as biomarkers for the diagnosis of mastitis in dairy cows.


Subject(s)
Gene Expression Profiling , Mammary Glands, Animal/metabolism , MicroRNAs/metabolism , Staphylococcus aureus/genetics , Animals , Biomarkers/metabolism , Cattle , DNA, Bacterial/chemistry , DNA, Bacterial/metabolism , Female , Gene Library , Mastitis, Bovine/diagnosis , Mastitis, Bovine/genetics , Mastitis, Bovine/microbiology , Models, Biological , Real-Time Polymerase Chain Reaction , Sequence Analysis, RNA , Staphylococcus aureus/isolation & purification
2.
Mol Neurobiol ; 50(3): 971-8, 2014 Dec.
Article in English | MEDLINE | ID: mdl-24788682

ABSTRACT

Heme oxygenase (HO) represents an intrinsic antiinflammatory system based on its ability to inhibit expression of proinflammatory cytokines. The constitutive isoform heme oxygenase-2 (HO-2) has high expression and activity in cerebral microvascular endothelial cells (CMVEC). This study was undertaken to evaluate the role of HO-2 in regulation of TLR4/MyD88-dependent signaling and to study the effect of HO-2 on the expression and secretion of the proinflammatory cytokines tumor necrosis factor α (TNF-α) and Interleukin-6 (IL6) in CMVEC. HO-2 short hairpin RNA (shRNA) and HO-2 overexpression plasmids were used to observe the effect of HO-2 on proinflammatory cytokines in CMVEC in vitro, and the results showed that the messenger RNA (mRNA) and protein levels of TNF-α and IL6 were increased and decreased, respectively, compared with control groups. LPS-stimulated TNF-α and IL6 mRNA and protein were also reduced in CMVEC treated with an inhibitor of TLR4 signaling, CLI-095, or HO-2 overexpression. CLI-095 and HO-2 overexpression both reduced TLR4 expression in CMVEC, and HO-2 shRNA blocked these effects of CLI-095. CLI-095 and HO-2 overexpression potently suppressed TLR4/MyD88-dependent proinflammatory cytokine expression in CMVEC. These results suggest that HO-2 plays an important role in protecting CMVEC against cytokine-mediated inflammation.


Subject(s)
Endothelial Cells/metabolism , Endothelium, Vascular/metabolism , Heme Oxygenase (Decyclizing)/metabolism , Interleukin-6/metabolism , Signal Transduction/physiology , Toll-Like Receptor 4/metabolism , Tumor Necrosis Factor-alpha/metabolism , Animals , Cells, Cultured , Cerebral Cortex/cytology , Cerebral Cortex/drug effects , Cerebral Cortex/metabolism , Endothelial Cells/cytology , Endothelial Cells/drug effects , Endothelium, Vascular/cytology , Endothelium, Vascular/drug effects , Heme Oxygenase (Decyclizing)/genetics , Interleukin-6/genetics , Lipopolysaccharides/pharmacology , Mice , RNA, Small Interfering , Signal Transduction/drug effects , Sulfonamides/pharmacology , Tumor Necrosis Factor-alpha/genetics
3.
Parasitol Int ; 63(2): 392-6, 2014 Apr.
Article in English | MEDLINE | ID: mdl-24412715

ABSTRACT

Hypodermin A (HA), a serine protease secreted by first-instar larvae of Hypoderma lineatum (Diptera: Oestridae) is associated with inflammatory and the specific immune responses in cattle hosts. In the present study, the cDNA sequence of HA was synthesized, and found to have fifteen amino acids which differed from the sequence available in GenBank. We then examined the association between recombinant HA and guinea-pig complement component 3 (C3) through a co-immunoprecipitation assay. Cos7 cells stably expressing HA were generated, and were found to be more resistant to lysis by guinea-pig C3 than the controls. HA was also able to degrade the C6 and C5b-9 of guinea-pig C3. The presumed DNA binding site of HA with guinea-pig C3 was detected by an electrophoretic mobility shift assay (EMSA). In contrast, after stable transfection, mHA was unable to reduce the amount of C3 or to inhibit its cytotoxicity, while HA could degrade guinea-pig C3 and inhibit the complement pathway. The findings suggest that recombinant HA could serve as an immunosuppressive agent against organ rejection after xenotransplantation.


Subject(s)
Complement System Proteins/metabolism , Serine Endopeptidases/pharmacology , Amino Acid Sequence , Animals , COS Cells , Chlorocebus aethiops , DNA, Complementary/genetics , Molecular Sequence Data
4.
Yi Chuan ; 32(12): 1256-62, 2010 Dec.
Article in Chinese | MEDLINE | ID: mdl-21513151

ABSTRACT

The polymorphism of Interleukin-8 (IL8) gene were investigated for 610 Chinese Holstein cows of 30 bull families from a dairy farm in Shanghai using Polymerase Chain Reaction-Single Strand Conformation Polymorphism (PCR-SSCP) technique with a mixed animal model to verify the effects of the polymorphisms on some milk productive performance, tested day milk yield, tested day fat percentage, tested day milk protein percentage, 305 d corrected milk yield, 305 d milk fat yield, 305 d milk protein yield, and somatic cell score (SCS). The aim was to explore the significant molecular marker in practical dairy production. Three genotypes were identified and the genotypic frequencies of KK, KA, and AA were 0.187, 0.451, and 0.362, respectively. The gene frequencies of K and A were 0.412 and 0.588. The results showed highly significant (P < 0.01) association of IL8 mutations with tested day milk yield, 305 d milk protein yield, 305 d corrected milk yield and 305 d milk fat yield, SCS and tested day milk protein percentage (P < 0.05). However, no association (P > 0.05) with tested day milk fat percentage was recorded. The cows with KK genotype had higher tested day milk yield, 305 d milk protein yield, 305 d corrected milk yield and 305 d milk fat yield than those with AA and KA genotypes (P < 0.01). The least square mean of SCS for KK was significantly lower than that with AA and KA genotypes (P < 0.01). AA genotype was significant lower in tested day milk protein percentage than KK and KA genotypes (P < 0.05). The IL8 gene genetic diversity has a great genetic effect on milk traits and mastitis resistance and could be a useful genetic marker for Chinese Holstein breeding.


Subject(s)
Cattle/genetics , Cattle/physiology , Interleukin-8/genetics , Lactation/genetics , Milk/metabolism , Polymorphism, Genetic/genetics , Animals , Base Sequence , Biomarkers/metabolism , Cattle/metabolism , China , Female , Gene Frequency , Genotype , Leukocytes/metabolism , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational
5.
Yi Chuan ; 30(1): 65-70, 2008 Jan.
Article in Chinese | MEDLINE | ID: mdl-18244904

ABSTRACT

The gayal (Bos frontalis) is a very rare, semi-wild and semi-domestic bovine species. There still exist remarkable divergences on the gayal's origin and phylogenetic status. The cytochrome b (Cyt b) gene entire sequences (1,140 bp) of 11 gayals were sequenced and analyzed. Combined with other bovine Cyt b entire sequences cited in GenBank, the phylogenetic trees of genus Bos were reconstructed by neighbor-joining (NJ) and maximum parsimony (MP) methods with Bubalus bubalis as outgroup. Sequence analysis showed that, among 1,140 sites of Cyt b gene entire sequences of 11 gayals, 95 variable sites (8.33% of all sites) and 6 haplotypes were found, showing abundant genetic diversity in mitochondrial Cyt b gene of the gayals. Both NJ and MP trees demonstrated that the gayals in this study were markedly divided into three embranchments: one embranchment clustering with Bos taurus, another clustering with Bos indicus, and the third clustering with Bos gaurus. The result of phylogenetic analysis suggested that the gayal might be the domesticated form of the gaur (Bos gaurus), and a great proportion of the gayal bloodline was invaded by other bovine species.


Subject(s)
Cytochromes b/genetics , Phylogeny , Ruminants/classification , Ruminants/genetics , Animals , Base Sequence , Databases, Genetic , Genetic Variation , Haplotypes , Molecular Sequence Data , Sequence Analysis, DNA
6.
Yi Chuan ; 29(3): 324-8, 2007 Mar.
Article in Chinese | MEDLINE | ID: mdl-17369154

ABSTRACT

Gene flow was analyzed among four sheep breeds of Mongolian Group in China using structural gene loci and microsatellite markers.Results showed that genetic differentiation was 0.0164-0.0455 by using structural loci, and was 0.0107-0.0239 by using microsatellite loci.This illustrated that most variations existed within breeds and genetic differentiation level was very low among sheep breeds of Mongolian Group in China.There was substantial Gene flow among the breeds as estimated by the structural loci (Nm=7.971) or the microsatellite method(Nm=15.732).There was no direct relationship between genetic differences from breeds and geographical distances.It is concluded that genetic differentiation of sheep breeds of Mongolian Group in China is mainly the result of natural selection (different living conditions).


Subject(s)
Gene Flow , Gene Frequency , Genetic Drift , Growth Hormone/genetics , Polymorphism, Genetic , Alleles , Animals , Breeding , China , Female , Genetics, Population , Male , Microsatellite Repeats/genetics , Polymerase Chain Reaction , Polymorphism, Single-Stranded Conformational , Sheep, Domestic
7.
Yi Chuan ; 29(2): 201-5, 2007 Feb.
Article in Chinese | MEDLINE | ID: mdl-17369177

ABSTRACT

Using the method of multiloci electrophoresis, genetic co-adaptability of Hu Sheep was analyzed according to allele frequencies in 14 structural loci. The results showed that no co-adaptability existed in the dominance-dominance model, while co-adaptability was observed in X-p-Cat combination locus of dominance-codominance model and in Po-CA, Po-Cat combination locus of codominance-codominance model. The findings demonstrated that co-adaptability occurred among neutral structural loci and played a key role in maintaining genetic situation, equilibrium or disequilibrium.


Subject(s)
Linkage Disequilibrium/genetics , Sheep/genetics , Alleles , Animals , Gene Frequency , Models, Genetic
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