利用单根肌纤维法分离和培养猪骨骼肌卫星细胞及其成肌诱导分化

建立单根肌纤维法体外培养猪骨骼肌卫星细胞的体系,了解其增殖和成肌特性。通过Ⅰ型胶原酶消化,从猪骨骼肌中分离完整的单根肌纤维并培养,用细胞免疫荧光鉴定肌纤维上的卫星细胞,随后对从单根肌纤维上游离出来的卫星细胞进行细胞免疫荧光染色,传代培养,成肌诱导分化和Western blot分析骨骼肌卫星细胞成肌特异性蛋白的表达。结果显示:分离并培养的单根肌纤维上附着有卵圆形的细胞,并随时间的推移,细胞缓慢向外迁移并增殖,卫星细胞特异性标志基因对盒转录因子(Paired protein box,Pax7)和成肌分化抗原(Myogenic Differentiation Antigen,MyoD)免疫荧光染色呈阳性,且阳性率达到90%以上。成肌诱导分化后,细胞开始汇合,并呈方向性生长,最终形成多核肌管,且成肌特异性标志基因Myogenin和myosin heavy chain(MyHC)表达呈阳性。MyoD蛋白高表达于增殖期,而Myogenin和MyHC在进入分化期才表达。该实验成功建立了猪骨骼肌单根肌纤维的体外培养方法并获得了高纯度的卫星细胞,为骨骼肌卫星细胞进行活体移植治疗相关疾病研究提供了实验材料。     

The aetiology of spinal deformity in Atlantic salmon, Salmo salar L.: influence of genetic factors on the frequency and severity in freshwater stages

A radiographic study was carried out on 2-year groups of pre-smolt parr to detect the comparative frequency of vertebral change in a range of defined pedigreed families of high growth rate of Atlantic salmon, Salmo salar L., reared under identical conditions. The observed frequency of deformity was related to the observed deformity level in the seawater parental generation, and to a potentially uncontrolled environmental determinant, i.e. stripping date. High and low parental deformity groups of families were found to produce offspring that had very similar levels of radiographically detectable spinal deformities in the pre-smolt parr, suggesting no genetic link. With regard to stripping date, the overall incidence was nearly half that seen in the preceding year (8.7% cf 16.6%). However, a non-significant trend to fewer radiographic spinal deformities was seen in the later stripping period.     

CORRECTION OF ANGULAR DEFORMITY BY ASYMMETRIC PHYSEAL GROWTH AND BONE REMODELLING

Angular deformities of 25 or 37 degrees valgus were created by transverse wedge osteotomies in the diaphysis of the radius in 3-month-old Cocker Spaniels. Spontaneous correction was judged from radiographs until skeletal maturity. The magnitude of correction was estimated, using the opposite normal leg as a control. Asymmetrical physeal growth corrected the induced angulation between the proximal and distal radial joint plane surfaces (joint plane angle). The average correction of the joint plane angle was 80% in the 25 degree group, and 72% in the 37 degree group. Bone remodeling of the radial shaft corrected the induced diaphyseal angulation by 50% in both groups during the study.     

Spinal deformity in triploid grass carp Ctenopharyngodon idella (Valenciennes)

From mid-2004 to mid-2005, several grass carp, Ctenophayngodon idella (Valenciennes), showing evidence of spinal deformity were presented to the Aquatic Animal Health Program, Cornell University. The carp were from three separate locations in New York State. The first case involved several fish from a natural body of water in the Catskill Mountain region of south-eastern New York State. The second was a single affected individual from a private pond in the Fingerlakes region of Central New York State. The third was a single individual from the Cold Springs Harbor Fish Hatchery, Cold Springs Harbor, Long Island. All fish were at least 7 years of age. Radiographs and computed tomography (CT) scans revealed the deformities to be of bony origin. The spinal deformities were characterized by variable amounts of kyphosis, scoliosis and rotation. While it is not possible to determine the specific cause(s) of the lesions, we consider a genetic component as a likely contributor to the observed pathology.     

3-甲基-4-硝基酚对雄性小鼠生殖毒性的研究

目的探讨3-甲基-4-硝基酚(PNMC)对雄性小鼠的生殖毒性作用。方法 20只小鼠随机分成阴性对照组和PNMC组(分0.9、9和90mg/kg·bw组),经口灌胃染毒30天,观察睾丸和精子的形态变化。结果染毒组小鼠各剂量睾丸生精细胞发生坏死,曲细精管腔内出现细胞团,精子细胞分化异常,形成均质红染颗粒,睾丸曲细精管生精上皮/直径比值在90mg/kg组明显高于对照组;染毒组小鼠精子畸形率明显高于对照组(P<0.05),并且随着染毒浓度的增加,畸形率有增加的趋势。结论 3-甲基-4-硝基酚不仅可以引起生精细胞坏死和抑制精子形成,还可以诱导精子畸变,从而对生殖系统造成损伤。     

Use of hinged circular fixator constructs for the correction of crural deformities in three dogs

Background Hinged circular external skeletal fixator constructs are used to perform sequential correction of angular limb deformities, often with resultant limb segment lengthening, via distraction osteogenesis. Although there are several reports describing the use of these constructs for correction of antebrachial deformities in dogs, there is little information regarding their use on other limb segments. This report describes the use of hinged circular fixator constructs for the correction of acquired crural deformities in three skeletally immature dogs. Case reports Two dogs had purely frontal plane deformities (one valgus, one varus) and the third dog had frontal (valgus) and sagittal (recurvatum) components to its deformity. At the time of long‐term evaluation, frontal plane angulation relative to the contralateral limb improved from 40° to 22° of valgus, 30° to 5° of valgus, and 20° to 1° of varus in the three individual dogs. Tibial length discrepancies of 12% and 22% that were initially present in two dogs were improved to 6% and 10%, respectively, of the contralateral tibial length at the time of final evaluation; both dogs had compensatory growth of the ipsilateral femur and all dogs had an excellent functional outcome. Conclusion These cases illustrate the value of using hinged circular fixator constructs for correction of crural angular deformities, particularly when length discrepancies of the tibia are present.     

Skeletal deformities associated with nutritional congenital rickets in newborn lambs

Abstract

CASE HISTORY: A group of 545 pregnant rising 2-year-old Coopdale ewes on a Southland sheep farm were grazed over winter on a fodder beet (Beta vulgaris) crop. Subsequently, 45 out of approximately 750 lambs were born with a variety of skeletal deformities, including shortened limbs, varus and valgus angular limb deformities, palmar grade stance and cranial bowing of the carpus. Analysis of the crop showed the fodder beet contained a low percentage of phosphorus. In addition, 60 out of 460 rising 2-year-old ewes that had been grazed on the fodder beet crop as 1-year-olds had incisor abnormalities and malocclusion.

PATHOLOGICAL FINDINGS: Two affected lambs (1-day-old and 3-days-old) with representative clinical signs examined postmortem were found to have markedly enlarged costochondral junctions, and noticeably enlarged long bone metaphyses. In addition, one lamb had a dense band of metaphyseal sclerosis beneath the physes of all long bones examined. Histopathological findings included small islands and columns of chondrocytes and eosinophilic cartilage matrix present in the metaphysis. Metaphyseal trabeculae were disorganised and often lined by accumulations of pale pink osteoid; similar pale pink osteoid was also present in the cortices. Unerupted molar teeth in the affected lambs lacked a layer of enamel, and the dentine was irregular with globular basophilia.

DIAGNOSIS: The gross and histopathological lesions were consistent with a diagnosis of rickets.

CLINICAL RELEVANCE: Nutritional congenital rickets has not been previously diagnosed in sheep, but is a recognised disease of human infants with vitamin D deficient mothers. The rickets in affected lambs was most likely associated with phosphorus deficiency as a result of the pregnant ewes grazing fodder beet during gestation. While vitamin D deficiency was not definitively ruled out in these cases, practitioners are alerted to the possible effects of feeding phosphorus-deficient fodder beet to ewes for long periods during gestation and to 1-year-old sheep during important growth periods.     

Unusual congenital deformities of the lower limb in two calves

Congenital deformities of the lower limb were observed in 2 calves from different properties. One was a newborn Holstein-Friesian bull calf with polydactyly and polypodia of the right fore-limb.The other was an 8-month-old Friesian/Jersey crossbred heifer calf, which had a mirror image duplication of the plantar/palmar half of the distal portion of the digits. The abnormalities in the second calf have not previously been described in cattle. This paper presents details of the clinical, radiological and necropsy findings in these 2 calves.     

藏绵羊不同部位骨骼肌营养成分、肌纤维特性及与相关基因表达的关联分析

旨在比较藏绵羊母羊7个部位肌肉（臂三头肌、冈上肌、股四头肌、背最长肌、腰大肌、胸斜方肌和小腿肌）的营养成分、肌纤维特性及其与产肉性能相关基因（ AKT1、 mTOR和 FABP3）表达水平的相关性。结果表明,藏绵羊不同部位肌肉含水量均在75%左右;小腿肌的蛋白含量最高,其次是背最长肌（P>0.05）,腰大肌的蛋白含量低于小腿肌（P<0.01）;背最长肌肌内脂肪（IMF）含量高于除腰大肌外的其他部位（P<0.05）,而小腿肌IMF含量最低（1.15%）。肌纤维横截面积大小为冈上肌>股四头肌>臂三头肌>小腿肌>胸斜方肌>腰大肌>背最长肌;肌纤维直径是股四头肌>冈上肌>小腿肌>臂三头肌>腰大肌>胸斜方肌>背最长肌;背最长肌密度最大,高于冈上肌（P<0.05）;肌纤维面积、直径与肌内脂肪含量均呈负相关,而肌纤维密度与IMF含量呈正相关;肌纤维面积与肌纤维直径呈极显著正相关（r=0.842）,而肌纤维面积和肌纤维直径与肌纤维密度呈极显著负相关（r=-0.906、-0.831）。 AKT1基因在小腿肌中的相对表达量高于其他部位（P<0.05）,而 mTOR基因在股四头肌中的相对表达量高于小腿肌、背最长肌（P>0.05）,显著高于臂三头肌、冈上肌及腰大肌（P<0.05）,与胸斜方肌差异极显著（P<0.01）; FABP3基因在小腿肌中的相对表达量低于除冈上肌外的其他部位（P<0.05）。 AKT1和 mTOR基因表达量与肌肉蛋白含量均呈正相关,其中 AKT1基因与蛋白含量呈极显著正相关（r=0.601）; FABP3基因表达量与IMF含量呈极显著正相关（r=0.728）。综合分析可知,藏绵羊小腿肌更符合人们对高蛋白、低脂肪羊肉特点的要求,而背最长肌相比于其他部位嫩度更好; AKT1和 mTOR基因表达量与藏绵羊不同部位肌肉蛋白质合成呈正相关, FABP3基因表达量与藏绵羊不同部位肌肉IMF沉积呈正相关。