Three-dimensional modelling of computed tomography images of limb bones in gazelles (Gazella subgutturosa)

The objective of the present study was to model in 3D the detailed morphometric features of the thoracic and pelvic limb bones of the gazelle (Gazella subgutturosa), whose natural habitat is only in the Sanliurfa province, by using multidetector computerized tomography (MDCT) images. Materials from 14 (7 male and 7 female) dead gazelles were collected from the Harran University Veterinary Medicine Faculty clinics during or after treatment. MDCT images of the gazelles were taken and saved as DICOM (Digital Imaging and Communications in Medicine) files. MIMICS 20.1 software was used to generate a 3D reconstruction of the data, and morphological and morphometrical evaluation was performed. In respect of morphological features, the combined metacarpus (Mc) III and metacarpus Mc IV were the main weight-bearing metacarpal bone, while Mc II and Mc V were rudimentary. On the femur, the lateral condyle was bigger than the medial condyle at the caudodistal end and the medial wing of the trochlea femoris was larger than the lateral. As with the thoracic limb, the combined metatarsus (Mt) III and metatarsus Mt IV were the main weight-bearing metacarpal bone and they displayed a synostosis structure. There was no secondary metatarsus in the gazelles’ pelvic limb. Based on digital measurements, the tibia was the longest bone of the gazelles’ skeletal system, and there was a statistically significant sexual dimorphism throughout the skeleton at the levels of p < .05, p < .01 and p < .001. As a result, the present study's findings are useful as reference data for further studies on anatomy, surgery and archaezoology, while also being useful for classifying species and for forensic sciences.     

维生素K_3对五龙鹅生长性能、屠宰性能及养分表观利用率的影响

本试验旨在研究饲粮中维生素K3添加水平对五龙鹅生长性能、屠宰性能及养分表观利用率的影响。试验分为2个阶段。1~4周龄阶段,选用1日龄五龙鹅360只,随机分为6个组,每个组6个重复,每个重复10只鹅。Ⅰ组为对照组,饲喂基础饲粮(维生素K3含量1.23 mg/kg),Ⅱ~Ⅵ组在基础饲粮中分别添加1、2、4、8、16 mg/kg的维生素K3。5~16周龄阶段,选用28日龄五龙鹅324只,随机分为6个组,每个组6个重复,每个重复9只鹅。A组为对照组,饲喂基础饲粮(维生素K3含量1.18 mg/kg),B~F组在基础饲粮中分别添加2、4、8、16、32 mg/kg的维生素K3。试验期16周。结果表明:1)经回归分析得出,1~4周龄阶段五龙鹅饲粮中添加4.81 mg/kg维生素K3,平均日增重最大;5~16周龄阶段饲粮中添加11.59 mg/kg维生素K3,平均日增重最大。2)与对照组相比,1~4周龄阶段,饲粮中添加4 mg/kg维生素K3能显著或极显著提高鹅的全净膛率和腹脂率(P0.05或P0.01);5~16周龄阶段,饲粮中添加8 mg/kg维生素K3能显著或极显著提高屠宰率、半净膛率、全净膛率和腿肌率(P0.05或P0.01)。3)与对照组相比,5~16周龄阶段,饲粮中添加8 mg/kg维生素K3能极显著提高钙的表观利用率(P0.01),显著提高干物质、粗蛋白质、粗脂肪、中性洗涤纤维、酸性洗涤纤维和磷的表观利用率(P0.05)。由此可见,本试验条件下,1~4周龄和5~16周龄五龙鹅饲粮中维生素K3适宜添加水平分别为4.81和11.59 mg/kg。     

DNA methylation and targeted sequencing of methyltransferases family genes in canine acute myeloid leukaemia,modelling human myeloid leukaemia

Tumours shows aberrant DNA methylation patterns, being hypermethylated or hypomethylated compared with normal tissues. In human acute myeloid leukaemia (hAML) mutations in DNA methyltransferase (DNMT3A) are associated to a more aggressive tumour behaviour. As AML is lethal in dogs, we defined global DNA methylation content, and screened the C‐terminal domain of DNMT3 family of genes for sequence variants in 39 canine acute myeloid leukaemia (cAML) cases. A heterogeneous pattern of DNA methylation was found among cAML samples, with subsets of cases being hypermethylated or hypomethylated compared with healthy controls; four recurrent single nucleotide variations (SNVs) were found in DNMT3L gene. Although SNVs were not directly correlated to whole genome DNA methylation levels, all hypomethylated cAML cases were homozygous for the deleterious mutation at p.Arg222Trp. This study contributes to understand genetic modifications of cAML, leading up to studies that will elucidate the role of methylome alterations in the pathogenesis of AML in dogs.     

RACE法扩增梅花鹿β-防御素-1(siBD-1)全长cDNA

为了获得梅花鹿β-防御素-1（sika deer β-defensin-1,siBD-1）cDNA全序列,本试验以梅花鹿舌黏膜组织内提取的总RNA为模板,根据前期已获得的siBD-1 cDNA的已知部分序列设计引物,采用5'-RACE和3'-RACE技术分别扩增5'-和3'-末端序列,将此扩增产物克隆入pMD18-T载体,进行PCR、双酶切鉴定及序列测定与分析。结果表明,成功克隆出长度约为172和299 bp的siBD-1 cDNA 5'-和3'-末端序列,从而得到418 bp的siBD-1 cDNA全序列（GenBank登录号：HM588696.1）,其中包含89 bp 5'-非翻译区（UTR）、192 bp的开放阅读框（ORF）、终止密码子TAA、118 bp的3'-UTR和Poly（A）16。同源性比对结果显示,siBD-1 cDNA与水牛的肠防御素（BEBD）同源性最高,为90.6%,与牛（EBD、LAP、TAP、BNBD-4）、山羊（GBD-1、GBD-2）、驯鹿（reBD-1）、绵羊（sBD-1、sBD-2）和骆驼（caBD-1）的防御素cDNA的同源性较高,分别为83.2%、83.1%、87.3%、87.0%、87.5%、87.5%、84.4%、79.9%、77.1%和70.5%;与马（hoBD-1）和猪（pBD-1）的同源性较低,为60.3%和72.4%;而与人（hBD-2）的同源性最低,为16.0%。siBD-1成熟肽由38个氨基酸残基组成,其中包含9个带正电荷的氨基酸残基。     

Prokaryotic Expression and Bioinformatics Analysis of Nonstructure Protein 3C from Porcine Encephalomyocarditis Virus

To obtain the 3C protein of encephalomypcarditis virus (EMCV) and predict its structure,3C gene was amplified by RT-PCR with the RNA as template from HB10 strain. And the target gene inserted into pMD18-T vector. The plasmids were sequenced after verification by PCR and double enzyme digestion. The target gene was cleaved from the correct plasmid,and inserted into the pET32a vector. The recombinant pET32a-EMCV-3C was transformed into TranSetta (DE3) and then induced by IPTG. Then the size was identified by SDS-PAGE and the antigenicity was analyzed by Western blotting. According to the sequence results,the length of 3C gene was 615 bp,which encoded 205 amino acids. The results of SDS-PAGE showed that His-3C protein mainly existed in the form of inclusion body with the molecular of 40 ku. The Western blotting results verified that purified His-3C could react with rabbit anti-EMCV serum prepared with EMCV. Bioinformatics analysis indicated that 3C protein was non-secretory,and had multiple phosphorylation sites,but it had no transmembrane region. Thus,the 3C protein mainly involved in protein hydrolysis process.3C protease,as the only protease in the EMCV genome,played an indispensable role in viral replication. In this study,we successfully constructed the prokaryotic expression vector of 3C protein and predicted its structure,which provided a basis for further study of the role of 3C protein and catalytic mechanism.     

Establishment of Digital RT-PCR Assay for Detection of Swine Influenza Virus H3N2 Subtype

A new assay for the detection of swine influenza virus (SIV) was developed with a novel nucleic acid probe——Molecular beacon in this study. The specific primers and molecular beacon probes were designed according to the conserved region of H3 and N2 genes of SIV H3N2 subtype. A digital RT-PCR assay was developed for detection of SIV H3N2 subtype. The results showed that SIV H3N2 subtype could be identified simultaneously on this microarry with high sensitivity and reproducibility,which could reach to 10⁶ dilute viruse. The conclusion was that the digital RT-PCR method could analyze quantitatively the RNA templates.On the identification of H3N2 SIV,the digital RT-PCR method was much more scientific than Real-time quantitative PCR method.     

Molecular Characterization of Equine Rotavirus Group A Detected in Argentinean Foals During 2009–2014

Equine rotavirus group A (RVA) has been detected in several countries worldwide since its first detection in 1975. Currently, equine RVA is considered the major cause of dehydrating diarrhea in foals younger than 3 months, and the frequency of detection in clinical cases varies from 20% to 77%. The genotypes of epidemiologic relevance found in horses are G3P[12] and G14P[12]. In a survey conducted in Argentina from 1992 to 2008, equine RVA was detected in 21% and 39% of the fecal samples and outbreaks, respectively. Genotype distribution was 51% G3P[12] and 33% G14P[12]. In continuation with the surveillance, the aim of the present study was to characterize the equine RVA detected in Thoroughbred foals in Argentina from 2009 to 2014. A total of 436 stool samples (corresponding to 177 single diarrhea cases or outbreaks) were analyzed. Equine RVA was detected in 31% (135 of 436) of the samples, which corresponded to 42% (74 of 177) of outbreaks. From the positive cases, 42% (57 of 135) were genotyped. Of this, 63% were G3 (36 of 57) and 37% (21 of 57) were G14 genotype. Considering the whole data (1992–2014), equine RVA was detected in 25% (300 of 1,207) of the stool samples and 41% (119 of 293) of the diarrhea outbreaks. The results of this study also show a cyclic pattern of the G3 and G14 prevalence in the horse population with a change in G3:G14 frequencies from year to year. Furthermore, clustering in the phylogenetic tree suggests evolutionary and geographic relationships between the Argentinean strains compared with the strain circulating worldwide.     

3-甲基喹噁啉-2-羧酸的制备及其细胞毒性

通过贝鲁特反应和还原、脱羧反应在体外合成制备3-甲基喹噁啉-2-羧酸,并进行理化鉴定和抗菌活性测试;通过MTT方法研究3-甲基喹噁啉-2-羧酸对多种细胞的生长抑制作用,利用单细胞电泳研究其对细胞DNA的损伤作用,利用流式细胞术研究其对细胞周期的改变。结果显示,体外成功合成制备出3-甲基喹噁啉-2-羧酸,而3-甲基喹噁啉-2-羧酸几乎没有抗菌效果,对多种细胞的生长抑制作用较弱,在剂量检测范围内细胞抑制率不到30%,但在一定剂量下能导致细胞DNA损伤,主要表现为尾长和尾部DNA含量显著升高;并且3-甲基喹噁啉-2-羧酸也能改变Chang细胞的细胞周期,表现为S期阻滞。结果表明,3-甲基喹噁啉-2-羧酸具有一定的细胞毒性。     

USE OF 99mTc-MERCAPTOACETYLTRIGLYCINE TO EVALUATE RENAL FUNCTION IN HORSES

Ten healthy horses were injected intravenously with 99mTc-MAG3 and the disappearance of radioactivity from the blood was measured. The total body clearance (Cl(B)) and elimination half-life (t1/2(beta)) were 7.9 +/- 1.5 ml/kg/minute and 32.8 +/- 4.1 minutes, respectively. The disappearance of 99mTc-MAG3 from the blood of 2 horses with compromised renal function was also measured. The data suggest that 99mTc-MAG3 is a useful and clinically applicable radiopharmaceutical for measurement of effective renal blood flow in the horse.