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1.
Environ Sci Pollut Res Int ; 30(19): 56641-56653, 2023 Apr.
Article in English | MEDLINE | ID: mdl-36920605

ABSTRACT

Alternative products such as those from high-value protien animals have increased the demand for the production of high-quality chicken meat in past few years. This study examines the impact of two distinct feeding types on goose body-weight, as well as the genetic variation of growth hormone (GH) and pituitary-specific transcription factor (Pit-1) genes in ten goose populations using single nucleotide polymorphism (SNP) markers and PCR-RFLP analysis. Both genes were seen as very important for productivity, especially in light of the COVID-19 and its effect on poultry industry at the time. The findings suggest that employing genetic indicators in these two genes in conjunction with a high-fat diet may be a feasible strategy for goose selection programme aiming to increase marketing body weight, as the high-fat diet outperformed the balanced diet. The study investigates the effect of gender, 2 types of diets, breeds and the genetic variation of the two genes, four SNPs were reported to be found: two at the GH gene exons C123T and C158T, and two at the Pit-1 gene exons G161A and T282G. Certain genotypes were found to have a substantial effect on the marketing body-weight of goose, which varied depending on the tested breeds. However, in terms of gender, males report higher and better performance levels than females. Diet, breeds and genotype interaction, and breeds, gender and genotype interaction were found to have a minor effect on goose body weight. However, diet, breeds, gender, SNP locus, diet and breeds interaction, and breeds and gender interaction were found to have a significant effect on goose body weight, as indicated by the effect size results.


Subject(s)
COVID-19 , Geese , Animals , Female , Male , Geese/genetics , Polymorphism, Single Nucleotide , Genotype , Transcription Factors/genetics , Body Weight , Meat
2.
Front Pediatr ; 10: 985878, 2022.
Article in English | MEDLINE | ID: mdl-36034570

ABSTRACT

Objective: To explore the potential of walking alone milestone combined reading-frame rule to improve the early diagnosis of Duchenne muscular dystrophy (DMD). Method: To retrospectively describe the genotype and phenotype of Duchenne and Becker muscular dystrophies (BMD) patients with deletions and duplicates in the dystrophin gene. The sensitivity and specificity of the reading frame rule were calculated and compared to that of the combined reading frame rule and walking alone milestone. The diagnostic coincidence rate of two different methods was analyzed. Result: One hundred sixty-nine male DMD/BMD patients were enrolled, including 17 cases of BMD and 152 cases of DMD. The diagnostic coincidence rate, diagnostic sensitivity, and specificity of the reading-frame rule for DMD/BMD were 85.2, 86.8, and 70.59%, respectively. The sensitivity and specificity of the reading frame principle combined with the walking alone milestone for DMD/BMD were 96.05 and 70.59%, respectively. The diagnostic coincidence rate increased to 93.49%, significantly different from that predicted by reading- frame rule (P < 0.05). Conclusion: The reading-frame rule combined with the walking alone milestone significantly improved the early diagnosis rate of DMD.

3.
Front Psychol ; 13: 898134, 2022.
Article in English | MEDLINE | ID: mdl-35664157

ABSTRACT

In the knowledge economy, the process of knowledge sharing and creation for value co-creation frequently emerge in a multi-agent and multi-level system. It's important to consider the roles, functions, and possible interactive knowledge-based activities of key actors for ecological development. Makerspace as an initial stage of incubated platform plays the central and crucial roles of resource orchestrators and platform supporter. Less literature analyses the knowledge ecosystem embedded by makerspaces and considers the interactive process of civil society and natural environment. This study constructs a multi-agent and multi-level knowledge ecosystem from macro, meso, and micro perspective based on Quintuple Helix theory and designs four evolutionary stages of knowledge orchestrating processes. This study finds that the symbiosis, co-evolution, interaction, and orchestration of multiple agents in the knowledge ecosystem should be merged with each other for value co-creation, which helps to take a systematic approach for policymakers, managers, and researchers.

4.
Sensors (Basel) ; 22(7)2022 Mar 23.
Article in English | MEDLINE | ID: mdl-35408070

ABSTRACT

Aiming at the demand for rapid detection of highway pavement damage, many deep learning methods based on convolutional neural networks (CNNs) have been developed. However, CNN methods with raw image data require a high-performance hardware configuration and cost machine time. To reduce machine time and to apply the detection methods in common scenarios, the CNN structure with preprocessed image data needs to be simplified. In this work, a detection method based on a CNN and the combination of the grayscale and histogram of oriented gradients (HOG) features is proposed. First, the Gamma correction was employed to highlight the grayscale distribution of the damage area, which compresses the space of normal pavement. The preprocessed image was then divided into several unit cells, whose grayscale and HOG were calculated, respectively. The grayscale and HOG of each unit cell were combined to construct the grayscale-weighted HOG (GHOG) feature patterns. These feature patterns were input to the CNN with a specific structure and parameters. The trained indices suggested that the performance of the GHOG-based method was significantly improved, compared with the traditional HOG-based method. Furthermore, the GHOG-feature-based CNN technique exhibited flexibility and effectiveness under the same accuracy, in comparison to those deep learning techniques that directly deal with raw data. Since the grayscale has a definite physical meaning, the present detection method possesses a potential application for the further detection of damage details in the future.


Subject(s)
Neural Networks, Computer
6.
Medicine (Baltimore) ; 99(43): e22940, 2020 Oct 23.
Article in English | MEDLINE | ID: mdl-33120854

ABSTRACT

RATIONALE: Acute encephalopathy with biphasic seizures and late reduced diffusion (AESD) has been reported almost exclusively in the Japanese population. PATIENT CONCERNS: A 17-month-old male patient presented with fever and seizures, and subsequently fell into a coma. On the second day, he recovered consciousness. On the fourth day, he developed complex partial seizures and fell into a coma again. On day 10, the fever and seizures subsided. Head computed tomography on the first day showed no abnormalities. Brain diffusion-weighted images on the fourth day revealed reduced diffusion in the bilateral subcortical white matter. DIAGNOSIS: A diagnosis of AESD was made. INTERVENTIONS: The patient was treated with corticosteroids and intravenous immunoglobulin. OUTCOMES: At the 4-month follow-up, the patient was able to walk independently, and the epileptic seizures were well controlled. LESSONS: AESD is a rare entity, and treatment with corticosteroids and intravenous immunoglobulin can lead to a favorable prognosis. Clinicians should be aware of this condition, and clinicoradiological features can suggest the diagnosis.


Subject(s)
Brain Diseases/complications , Diffusion Magnetic Resonance Imaging/methods , Seizures/etiology , Acute Disease , Adrenal Cortex Hormones/therapeutic use , Brain Diseases/diagnosis , Brain Diseases/drug therapy , Coma/diagnosis , Coma/etiology , Drug Therapy, Combination , Fever/diagnosis , Fever/etiology , Humans , Immunoglobulins, Intravenous/therapeutic use , Infant , Male , Seizures/diagnosis , Seizures/physiopathology , Tomography, X-Ray Computed/methods , Treatment Outcome , White Matter/diagnostic imaging , White Matter/pathology
7.
Front Genet ; 11: 46, 2020.
Article in English | MEDLINE | ID: mdl-32117456

ABSTRACT

DNA methylation is an epigenetic modification primarily responsible for individual phenotypic variation. This modification has been reported to play an important role in caste, brain plasticity, and body development in honeybees (Apis mellifera). Here, we report the DNA methylation profile of honeybee hypopharyngeal glands, from atrophy in winter to arousal in the following spring, through the use of whole-genome bisulfite sequencing. Consistent with previous studies in other Apis species, we found low methylation levels of the hypopharyngeal gland genome that were mostly of the CG type. Notably, we observed a strong preference for CpG methylation, which was localized in promoters and exon regions. This result further indicated that, in honeybees, DNA methylation may regulate gene expression by mediating alternative splicing, in addition to silencing gene in the promoter regions. After assessment by correlation analysis, we identified seven candidate proteins encoded by differentially methylated genes, including aristaless-related homeobox, forkhead box protein O, headcase, alpha-amylase, neural-cadherin, epidermal growth factor receptor, and aquaporin, which are reported to be involved in cell growth, proliferation, and differentiation. Hypomethylation followed by upregulated expression of these candidates suggested that DNA methylation may play significant roles in the activation of hypopharyngeal glands in overwintering honeybees. Overall, this study elucidates epigenetic modification differences in honeybee hypopharyngeal glands by comparing an inactive winter state to an aroused state in the following spring, which could provide further insight into the evolution of insect sociality and regulatory plasticity.

8.
Yi Chuan ; 41(10): 974-978, 2019 Oct 20.
Article in Chinese | MEDLINE | ID: mdl-31624059

ABSTRACT

"Animal Genetics Principles and Breeding Methods" is a main course for Master students majoring in Agriculture (Livestock) and involves a combination of theory and practice. The traditional teaching method is difficult not only to meet the requirements of modern professional degree teaching, but also for the students to master the theory and practice of genetic breeding. We have employed the case study methodology during the entire course. This paper analyzes the connotation and characteristics of the method and expounds the design and discussion of the cases. Besides, the teaching evaluation is also included. It provides a reference for the application and promotion of the case teaching method in training graduate students majoring in agriculture.


Subject(s)
Animal Husbandry/education , Breeding , Curriculum , Animals , Teaching
9.
Asian-Australas J Anim Sci ; 32(11): 1664-1672, 2019 11 01.
Article in English | MEDLINE | ID: mdl-30744373

ABSTRACT

OBJECTIVE: This study aimed to measure genetic diversity and to determine the relationships among fourteen goose breeds. METHODS: microsatellite markers were isolated from the genomic DNA of geese based on previous literature. The DNA segments, including short tandem repeats, were tested for their diversity among fourteen populations of geese. The diversity was tested on both breeds and loci level and by mean of unweighted pair group method with arithmetic mean (UPGMA) and structure program, phylogenetic tree and population structure were tested. RESULTS: A total of 108 distinct alleles (1%) were observed across the fourteen breeds, with 36 out of the 108 alleles (33.2%) being unique to only one breed. Genetic parameters were measured per the 14 breeds and the 9 loci. Medium to high heterozygosity was reported with high effective numbers of alleles (Ne). Polymorphic Information Contents (PIC) of the screened loci was found to be highly polymorphic for eleven breeds; while 3 breeds were reported moderately polymorphic. Breeding coefficient (FIS) ranged from -0.033 to 0.358, and the pair wise genetic differentiation (FST) ranged from 0.01 to 0.36 across the fourteen breeds; for the 9 loci Ho, He and Ne were same as the breeds parameters, PIC of the screened loci reported 6 loci highly polymorphic and 3 loci to be medium polymorphic, and FIS ranged from -0.113 to 0.368. In addition, genetic distance estimate revealed a close genetic distance between Can. and Hort. breeds by 0.04, and the highest distance was between Tai. and Gray. breed by 0.54. CONCLUSION: Cluster analyses were made, and they revealed that goose breeds had hybridized frequently, resulting in a loss of genetic distinctiveness for some breeds.

10.
CNS Neurosci Ther ; 25(1): 112-122, 2019 01.
Article in English | MEDLINE | ID: mdl-29808547

ABSTRACT

OBJECTIVE: To explore the role of miR-181b in alterations of apoptosis and autophagy in the kainic acid (KA)-induced epileptic juvenile rats via modulating TLR4 and P38/JNK signaling pathway. METHODS: Dual-luciferase reporter assay was performed to testify the targeting relationship between miR-181b and TLR4. After intracerebroventricular injection (i.c.v.) of KA, rats were injected with miR-181b agomir and TLR4 inhibitor (TAK-242). The TLR-4 activator lipopolysaccharide (LPS) was also administered into rats immediately after injection with miR-181b agomir. Quantitative real-time-polymerase chain reaction (qRT-PCR) was used for detections of miR-181b and TLR4 expressions, hematoxylin-eosin (HE) and Nissl staining for observation of the hippocampus morphological changes, and TUNEL staining for apoptosis analysis. Moreover, western blot was determined to detect TLR4 and P38/JNK pathway proteins, as well as autophagy- and apoptosis-related proteins. RESULTS: TLR4 was identified as a direct target of miR-181b using Dual-luciferase reporter assay. KA rats injected with miR-181b agomir or TAK-242 had improved learning and memory abilities, reduced seizure severity of Racine's scale, and lessened neuron injury. Additionally, miR-181b agomir or TAK-242 could significantly inhibit P38/JNK signaling, decrease LC3II/I, Beclin-1, ATG5, ATG7, ATG12, Bax, and cleaved caspases-3, but increase p62 and Bcl-2 expression. No significances were found between KA group and KA + miR-181b + LPS group. CONCLUSION: MiR-181b could inhibit P38/JNK signaling pathway via targeting TLR4, thereby exerting protective roles in attenuating autophagy and apoptosis of KA-induced epileptic juvenile rats.


Subject(s)
Apoptosis/physiology , Autophagy/physiology , Epilepsy/metabolism , MAP Kinase Signaling System/physiology , MicroRNAs/metabolism , Toll-Like Receptor 4/metabolism , Animals , Disease Models, Animal , Epilepsy/pathology , Female , Hippocampus/metabolism , Hippocampus/pathology , Kainic Acid , MAP Kinase Kinase 4/metabolism , Male , Neuroprotection/physiology , Random Allocation , Rats, Wistar , p38 Mitogen-Activated Protein Kinases/metabolism
11.
J Cell Physiol ; 234(7): 11330-11347, 2019 07.
Article in English | MEDLINE | ID: mdl-30478915

ABSTRACT

Salmonella Enteritidis (SE) can be transmitted to eggs through cecum or the ovary from infected layers and causes food poisoning in humans. The mechanism of cecal transmission has been extensively studied. However, the mechanism and route of transovarian transmission of SE remain unclear. In this study, the ducks were orally inoculated with SE, and the ovarian follicles and stroma were collected to detect SE infection. The immune responses were triggered and the innate and adaptive immune genes (TLR4, NOD1, AvßD7, and IL-1ß) were upregulated significantly during the SE challenge. Moreover, the ovary tissues (small follicle and stroma) of susceptible and resistant-laying ducks were performed by RNA sequencing. We obtained and identified 23 differentially expressed genes (DEGs) between susceptible and resistant-laying ducks in both small follicle and stroma tissues ( p < 0.05). The DEGs were predominately identified in the p53 signaling pathway. The expression of key genes (p53, MDM2, PERP, caspase-3, and Bcl-2) involved in the signaling pathway was significantly higher in granulosa cells (dGCs) from SE-infected ducks than those from uninfected ducks. Moreover, the overexpression of PERP resulted in further induction of p53, MDM2, caspase-3, and Bcl-2 during SE infection in dGCs. Whereas, an opposite trend was observed with the knockdown of PERP. Besides, it is further revealed that the PERP could enhance cell apoptosis, SE adhesion, and SE invasion in SE-infected dGCs overexpression. Altogether, our results demonstrate the duck PERP involved in the ovarian local immune niche through p53 signaling pathway in dGCs challenged with SE.


Subject(s)
Ducks/immunology , Ducks/microbiology , Gene Expression Profiling/veterinary , Granulosa Cells/metabolism , Membrane Proteins/metabolism , Salmonella Infections, Animal/immunology , Animals , Apoptosis/physiology , Cell Proliferation/physiology , Female , Gene Expression Regulation/genetics , Membrane Proteins/genetics , Membrane Proteins/immunology , Ovarian Follicle/immunology , Ovarian Follicle/microbiology , Salmonella Infections, Animal/microbiology , Salmonella Infections, Animal/pathology , Salmonella enteritidis/immunology , Tumor Suppressor Protein p53/metabolism
12.
Front Genet ; 9: 426, 2018.
Article in English | MEDLINE | ID: mdl-30349555

ABSTRACT

There are increasing concerns regarding the impact of agrochemical pesticides on non-target organisms. Pesticides could cause honeybee abnormal development in response to neurotoxins such as neonicotinoid. However, knowledge of carbendazim, a widespread fungicide in beekeeping practice, influencing on honeybee (Apis mellifera L.) brain development is lacking. Large-scale transcriptome approaches were applied to determine the changes in global gene expression in the brains of newly emerged honeybees after carbendazim exposure during the larval stage. To further understand the effects of carbendazim on the brain development of honeybees, the functions of differentially expressed genes were compared between the treatment and control groups. We found that neuroregulatory genes were down-regulated after carbendazim exposure, which suggest the neurotoxic effects of this fungicide on honeybee nervous system. Carbendazim exposure also altered the expression of genes implicated in metabolism, transport, sensor, and hormone. Notably, larvae in the carbendazim-treated group observed longer time to shift into the dormant pupal state than the control group. Moreover, a low juvenile hormone and high ecdysone titers were found in the treatment group compared to control group. The data is the first report of neurotoxic effects on honeybee caused by carbendazim, and the sublethal carbendazim may disturb honeybee development and is a potential chemical threating the honeybee colonies.

13.
PLoS One ; 12(8): e0183409, 2017.
Article in English | MEDLINE | ID: mdl-28837587

ABSTRACT

OBJECTIVE: To analyze the midterm evaluation data from the National Syphilis Prevention and Control Plan (2010-2020) and evaluate the current status of syphilis prevention and control in Jiangsu province, China. METHODS: We collected data via (1) field surveys conducted in 2015 and (2) data recorded in existing syphilis surveillance systems. We conducted descriptive statistical analysis to evaluate the current landscape of syphilis control initiatives and their potential effect in syphilis control. RESULTS: The incidence of all cases of syphilis decreased from 2010 (32.3 per 100,000) to 2015 (30.1 per 100,000), with an annual growth of -1.17% (x2trend = -7.52, P<0.001) in Jiangsu province. The incidence of primary and secondary syphilis and congenital syphilis both decreased significantly from 2010 to 2015. The average awareness rate of syphilis knowledge among professional personnel was 95.4% (3781/3963). Rural residents had the lowest awareness rate (83.5%, 1875/2245) and commercial sex workers had the highest awareness rate (92.1%, 7804/8474) in 2015. Only 47.8% (33908/70894) of patients received provider-initiated syphilis counseling and testing (PISTC) services in sexually transmitted disease (STD) clinics, but 94.5% (87927/93020) of all syphilis patients received free testing for syphilis. Overall, 97.2% (9378/9648) of syphilis reported cases of syphilis at medical institutions were confirmed to be accurate, and 92.2% (5850/6345) of patients diagnosed with syphilis at medical institutions received treatment with penicillin. CONCLUSION: The syphilis incidence rate in Jiangsu has decreased in recent years, but remains at a high level. It is essential to promote PISTC services to improve knowledge of syphilis and rates of testing and treatment in Jiangsu province.


Subject(s)
Syphilis/prevention & control , China/epidemiology , Cross-Sectional Studies , Humans , Incidence , Population Surveillance , Prevalence , Syphilis/epidemiology
14.
Mitochondrial DNA A DNA Mapp Seq Anal ; 27(6): 4588-4589, 2016 11.
Article in English | MEDLINE | ID: mdl-27210615

ABSTRACT

The bar-headed goose is known one of the world's highest-flying birds. In this study, the complete mitochondrial genome of Anser indicus (16,728 bp in length) was sequenced. Similar to the typical mtDNA of other vertebrates, goose mtDNA contained 37 genes (13 protein-coding genes, 2 rRNA genes and 22 tRNA genes) and a non-coding region (D-loop). The characteristics of the mitochondrial genome were analyzed in detail. We deduce that ND5 may be a major gene required for adaptation to high-altitude flight. The complete mitochondrial genome sequence of A. indicus obtained will be useful for phylogenetics, and biological characteristics.


Subject(s)
Anseriformes/genetics , Genome, Mitochondrial , Animals , Avian Proteins/chemistry , Avian Proteins/genetics , Avian Proteins/metabolism , Base Composition , Codon, Initiator , Codon, Terminator , DNA, Mitochondrial/chemistry , DNA, Mitochondrial/isolation & purification , DNA, Mitochondrial/metabolism , Open Reading Frames/genetics , RNA, Ribosomal/chemistry , RNA, Ribosomal/genetics , RNA, Transfer/chemistry , RNA, Transfer/genetics , Sequence Analysis, DNA
15.
Neurol India ; 64(2): 233-6, 2016.
Article in English | MEDLINE | ID: mdl-26954798

ABSTRACT

BACKGROUND: Intractable epilepsy is defined as the occurrence of seizures that cannot be controlled with medical treatment. The discovery of epilepsy biomarkers is increasingly attracting more attention from both clinical physicians as well as neuroscientists. Increased levels of soluble and/or cellular galectin-3 (Gal-3) have been associated with various diseases. However, the effects of Gal-3 in epilepsy are still unknown. In this study, we evaluated the association of higher interictal serum Gal-3 protein levels in patients diagnosed with intractable epilepsy. PATIENTS AND METHODS: A group of 38 patients with intractable epilepsy and 26 healthy age-matched control subjects were included in this study. A commercially available electrochemiluminescence immunoassay (ECLIA) kit was used to determine serum Gal-3 protein levels. RESULTS: Our results indicated that serum Gal-3 protein level in the patient group was 6.67 ± 0.34 ng/ml, and in the age-matched control group was 5.40 ± 0.34 ng/ml. The difference between the two groups was found to be statistically significant (P = 0.003). CONCLUSION: This study found a detectable elevation in serum Gal-3 concentration in patients with focal epilepsy. Given its secretory nature and detectable levels in the serum, Gal-3 could be a potential biomarker for intractable epilepsy.


Subject(s)
Drug Resistant Epilepsy/metabolism , Galectin 3/metabolism , Biomarkers/metabolism , Blood Proteins , Galectins , Humans
16.
J Steroid Biochem Mol Biol ; 152: 25-33, 2015 Aug.
Article in English | MEDLINE | ID: mdl-25864627

ABSTRACT

The hormonally active form of vitamin D3, 1α,25-(OH)2D3, has an important role in bone metabolism. This study examined the effects of 1α,25-(OH)2D3 on the ability of two cytokines, receptor activator of nuclear factor-κB ligand (RANKL) and macrophage-colony stimulating factor (M-CSF), to induce RAW 264.7 cells to form osteoclasts. A TRAP histochemical staining assay and bone resorption analysis were used to identify the rate of formation and activity of osteoclasts. The numbers of osteoclasts formed, and their bone resorption activity, was enhanced by the addition of 1α,25-(OH)2D3. The expression levels of osteoclast-specific proteins that are essential for bone resorption, integrin ß3, V-ATPase, CAII, CTSK, TRAP and MMP-9, were detected by western blotting. During 48 h, the expression levels of all these proteins significantly increased. Quantitative real-time polymerase chain reaction was used to determine the expression levels of the transcription factors, c-Fos and NFATcl. The expression levels of c-Fos and NFATc1 also increased 24h after treatment with 1α,25-(OH)2D3. These results suggest that 1α,25-(OH)2D3 can regulate bone metabolism by directly enhancing the formation and maturation of osteoclasts.


Subject(s)
Bone Resorption/metabolism , Cholecalciferol/pharmacology , Macrophage Colony-Stimulating Factor/metabolism , Osteoclasts/cytology , RANK Ligand/metabolism , Animals , Bone and Bones/metabolism , Cathepsin K/biosynthesis , Cattle , Cell Differentiation/drug effects , Cell Line , Cholecalciferol/analogs & derivatives , Integrin beta3/biosynthesis , Matrix Metalloproteinase 9/biosynthesis , Mediator Complex/biosynthesis , Mice , NFATC Transcription Factors/biosynthesis , Proto-Oncogene Proteins c-fos/biosynthesis , Vacuolar Proton-Translocating ATPases/biosynthesis
17.
Exp Ther Med ; 9(3): 909-912, 2015 Mar.
Article in English | MEDLINE | ID: mdl-25667652

ABSTRACT

Paroxysmal kinesigenic dyskinesia (PKD) is an autosomal dominant disorder and PRRT2 is the causative gene of PKD. The aim of this study was to investigate PRRT2 mutations in patients who were clinically diagnosed with PKD. Nine PKD cases, including four familial cases and five sporadic cases, were selected. Peripheral blood was drawn after obtaining informed consent, and genomic DNA was extracted by a standard protocol. Sanger sequencing was performed for the screening of PRRT2 mutations. A total of five cases were detected to harbor PRRT2 mutations. Four familial cases carried a c.649dupC (p.Arg217Profs*8) mutation, while one sporadic case and his asymptomatic father carried a c.133-136delCCAG (p.Pro45Argfs*44) mutation. PRRT2 mutations were not identified in the remaining cases. The study further confirmed that PRRT2 was a causative gene of PKD and implied that PRRT2 mutation has incomplete penetrance.

18.
Mol Biol Rep ; 42(2): 431-9, 2015 Feb.
Article in English | MEDLINE | ID: mdl-25332128

ABSTRACT

Cluster of differentiation 8 alpha (CD8α) is critical for cell-mediated immune defense and T-cell development. Although CD8α sequences have been reported for several species, very little is known about CD8α in ducks. To elucidate the mechanisms involved in the innate and adaptive immune responses of ducks, we cloned CD8α coding sequences from domestic, Muscovy, Mallard, and Spotbill ducks using reverse transcription polymerase chain reaction (RT-PCR). Each sequence consisted of 714 nucleotides and encoded a signal peptide, an IgV-like domain, a stalk region, a transmembrane region, and a cytoplasmic tail. We identified 58 nucleotide differences and 37 amino acid differences among the four types of duck; of these, 53 nucleotide and 33 amino acid differences were between Muscovy ducks and the other duck species. The CD8α cDNA sequence from domestic duck consisted of a 61-nucleotide 5' untranslated region (UTR), a 714-nucleotide open reading frame, and an 849-nucleotide 3' UTR. Multiple sequence alignments showed that the amino acid sequence of CD8α is conserved in vertebrates. RT-PCR revealed that expression of CD8α mRNA of domestic ducks was highest in the thymus and very low in the kidney, cerebrum, cerebellum, and muscle. Immunohistochemical analyses detected CD8α on the splenic corpuscle and periarterial lymphatic sheath of the spleen. CD8α mRNA in domestic ducklings was initially up-regulated, and then down-regulated, in the thymus, spleen, and liver after treatment with duck hepatitis virus type I (DHV-1) or the immunostimulant polyriboinosinic polyribocytidylic acid (poly I:C).


Subject(s)
CD8 Antigens/genetics , Ducks/genetics , Amino Acid Sequence , Animals , CD8 Antigens/metabolism , Cloning, Molecular , DNA, Complementary , Ducks/virology , Gene Expression , Hepatitis Virus, Duck , Organ Specificity/genetics , Phylogeny , RNA, Messenger/genetics , Sequence Alignment , Sequence Analysis, DNA
19.
Zhongguo Dang Dai Er Ke Za Zhi ; 16(12): 1215-9, 2014 Dec.
Article in Chinese | MEDLINE | ID: mdl-25523568

ABSTRACT

OBJECTIVE: To study the alterations of follicular T helper cells (CD4(+)CXCR5(+)Tfh cells, Tfh) on circulating T lymphocytes in children with asthma, and to study the expression of transcription regulatory factors BCL-6 and BLIMP-1 mRNA. METHODS: Sixty-four children with asthma and 25 healthy controls were enrolled in this study. On the basis of the disease, the children with asthma were classified into acute phase group (n=36) and remission phase group (n=28). The flow cytometry was used to detect the proportion of CD4(+)CXCR5(+)Tfh cells on CD4(+)T lymphocytes. Real-time PCR was performed to detect the levels of BCL-6 mRNA and BLIMP-1 mRNA. The double -antibody Sandwich ELISA was used to detect plasma concentrations of total IgE, IL-2, IL-6 and IL-21. RESULTS: The proportion of CD4(+)CXCR5(+)Tfh cells was significantly higher in the acute group than in the control group and the remission group (P<0.05). Transcription levels of BCL-6 mRNA were significantly higher, while the inhibitory factors BLIMP-1 mRNA was significantly lower in the acute group than in the remission group and control group (P<0.05). The plasma concentration of IL-6 in the acute group increased significantly compared with the control group (P<0.05). Plasma concentrations of total IgE and IL-21 increased significantly, in contrast, plasma IL-2 concentration decreased significantly in the acute group, compared with the control group and the remission group (P<0.05). Correlation analysis showed that both IL-21 and IL-6 concentrations were positively correlated with the proportion of CD4(+)CXCR5(+)Tfh cells (r=0.76, r=0.46 respectively; P<0.05), while IL-2 level was negatively correlated with the proportion of Tfh cells (r=-0.68, P<0.05). CONCLUSIONS: The abnormal proportion of CD4(+)CXCR5(+)Tfh cells might be involved in the immunological pathogenesis of acute asthma in children. The increased expression of BCL-6 mRNA and decreased expression of BLIMP-1 mRNA as well as the alterations of plasma total IgE, cytokines IL-2, IL-6 and IL-21 in microenvironment might be account for the increased proportion of CD4(+)CXCR5(+)Tfh cells in children with acute asthma.


Subject(s)
Asthma/immunology , DNA-Binding Proteins/genetics , Receptors, CXCR5/analysis , Repressor Proteins/genetics , T-Lymphocytes, Helper-Inducer/immunology , Child , Child, Preschool , Female , Humans , Immunoglobulin E/blood , Infant , Interleukins/blood , Male , Positive Regulatory Domain I-Binding Factor 1 , Proto-Oncogene Proteins c-bcl-6 , RNA, Messenger/analysis
20.
Mol Biol Rep ; 41(9): 6233-40, 2014 Sep.
Article in English | MEDLINE | ID: mdl-24981929

ABSTRACT

H-ferritin is a core subunit of the iron storage protein ferritin, and is related to the pathogenesis of malignant diseases. A differential expressed sequence tag of the ferritin, heavy polypeptide 1 gene (FTH1) was obtained from our previously constructed suppression subtractive cDNA library from 3-day-old ducklings challenged with duck hepatitis virus type I (DHV-1). The expression and function of FTH1 in immune defense against infection remains largely unknown in ducks. In this study, the full-length duFTH1 cDNA was obtained using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. It consisted of 153 basepairs (bp) 5'untranslated region (UTR), 183 bp 3'UTR, and 546 bp open reading frame that encodes a single protein of 181 amino acid residues. duFTH1 shares high similarity with FTH1 genes from other vertebrates. The amino acid sequence possesses the conserved domain of typical ferritin H subunits, including seven metal ligands in the ferroxidase center, one iron binding region signature, and a potential bio-mineralization residue (Thy(29)). Moreover, in agreement with a previously reported ferritin H subunit, we identified an iron response element in the 5'UTR. RT-PCR analyses revealed duFTH1 mRNA is widely expressed in various tissues. Real-time quantitative polymerase chain reaction analyses suggested that duFTH1 mRNA is significantly up-regulated in the liver after DHV-1 injection or polyriboinosinic polyribocytidylic acid (polyI:C) treatment, reaching a peak 4 h post-infection, and dropping progressively and returning to normal after 24 h. Our findings suggest that duFTH1 functions as an iron chelating protein subunit in duck and contributes to the innate immune responses against viral infections.


Subject(s)
Apoferritins/genetics , Ducks/genetics , Amino Acid Sequence , Animals , Apoferritins/metabolism , Base Sequence , Cloning, Molecular , DNA, Complementary/genetics , Ducks/virology , Gene Library , Hepatitis Virus, Duck/isolation & purification , Hepatitis, Viral, Animal/drug therapy , Hepatitis, Viral, Animal/immunology , Iron/metabolism , Iron Chelating Agents/metabolism , Molecular Sequence Data , Nucleic Acid Conformation , Open Reading Frames , Phylogeny , Picornaviridae Infections/drug therapy , Picornaviridae Infections/immunology , Poly I-C/pharmacology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Response Elements/genetics , Sequence Analysis, DNA , Up-Regulation
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