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1.
Intensity‐modulated radiation therapy (IMRT) allows optimization of radiation dose delivery to complex tumor volumes with rapid dose drop‐off to surrounding normal tissues. A prospective study was performed to evaluate the concept of conformal avoidance using IMRT in canine sinonasal cancer. The potential of IMRT to improve clinical outcome with respect to acute and late ocular toxicity was evaluated. Thirty‐one dogs with sinonasal cancer were treated definitively with IMRT using helical tomotherapy and/or dynamic multileaf collimator (DMLC) delivery. Ocular toxicity was evaluated prospectively and compared with a comparable group of historical controls treated with conventional two‐dimensional radiotherapy (2D‐RT) techniques. Treatment plans were devised for each dog using helical tomotherapy and DMLC that achieved the target dose to the planning treatment volume and limited critical normal tissues to the prescribed dose–volume constraints. Overall acute and late toxicities were limited and minor, detectable by an experienced observer. This was in contrast to the profound ocular morbidity observed in the historical control group treated with 2D‐RT. Overall median survival for IMRT‐treated and 2D‐treated dogs was 420 and 411 days, respectively. Compared with conventional techniques, IMRT reduced dose delivered to eyes and resulted in bilateral ocular sparing in the dogs reported herein. These data provide proof‐of‐principle that conformal avoidance radiotherapy can be delivered through high conformity IMRT, resulting in decreased normal tissue toxicity as compared with historical controls treated with 2D‐RT.  相似文献   

2.
Proteases play important roles in modulating a wide range of cellular functions, in the regulation of biologic processes, and in the pathogenesis of various diseases. Several molecular techniques are available to identify and characterize proteases in cells and tissues. Most of these techniques do not provide information on the activity of proteases in tissues. In situ zymography (ISZ) is a relatively low-cost technique that uses specific protease substrates to detect and localize specific protease activities in tissue sections. Used in combination with other techniques, ISZ provides data that further our understanding of the role of specific proteases in various pathologic and physiologic conditions. This review describes the general principle of ISZ and highlights the past and future applications of this technique in molecular pathology.  相似文献   

3.
Differences in dose homogeneity and irradiated volumes of target and surrounding normal tissues between 3D conformal radiation treatment planning and simulated non‐graphic manual treatment planning were evaluated in 18 dogs with apocrine gland adenocarcinoma of the anal sac. Overall, 3D conformal treatment planning resulted in more homogenous dose distribution to target tissues with lower hot spots and dose ranges. Dose homogeneity and guarantee of not under‐dosing target tissues with 3D conformal planning came at the cost, however, of delivering greater mean doses of radiation and of irradiating greater volumes of surrounding normal tissue structures.  相似文献   

4.
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5.
为探究miR-107在静原鸡肌肉组织肌苷酸特异性沉积过程中的调控机制,本研究利用相关生物信息学软件和数据库对静原鸡miR-107进行了靶基因预测与生物信息学分析。通过转录组测序获取静原鸡miR-107序列,进行序列比对和保守性分析;通过TargetScan、miRDB和PicTar预测结果的交集并结合miRTarbase中已证实的靶基因集合,取二者并集,采用OmicShare工具对基因集合进行GO功能富集和信号通路富集分析,并讨论HMED数据库中miR-107在不同组织和疾病中的表达水平。结果显示,miR-107序列在各物种间高度保守,miR-107调控的靶基因GO功能富集主要包括细胞过程、生物调节、代谢过程、刺激反应等生物学过程和结合、催化活性、转录调节活性等分子功能;靶基因存在于细胞的多个组分中,包括细胞器、细胞膜、含蛋白质复合物。信号转导通路显著富集于癌症中的microRNAs途径（microRNAs in cancer）、人乳头瘤病毒感染（human papillomavirus infection）、癌症中途径（pathways in cancer）、PI3K-Akt信号通路（PI3K-Akt signaling pathway）中（P<0.05）。miRNA-靶基因互作分析发现,miR-107主要通过与靶基因mRNA的非翻译区靶向结合抑制靶基因的表达。根据HMED数据库中miR-107在不同组织和疾病中的表达数据,发现miR-107在黑素瘤（melanoma）中的表达水平最高,在扁桃体（tonsil）和肌肉（muscle）等组织中表达水平较低。通过对静原鸡miR-107靶基因的生物信息学分析,为miR-107功能及调控机制的深入研究提供参考依据,也为进一步研究miR-107在静原鸡肌肉发育中的作用奠定基础。  相似文献   

6.
Sheep have been used as translational models of human postnatal testicular development. However, the morphometric features of the normal developing testis in sheep embryos have not been previously investigated using stereology. The objective of the present work was to establish normal quantitative parameters for fetal testicular tissue components in sheep, using unbiased design-based stereological methods. Twenty-four sheep embryos were divided into four gestational age groups (9–11, 12–14, 15–17 and 18–20 weeks of gestation) on the basis of the embryos’ crown-rump length. Isotropic, systematic uniform random sections of the left testes were obtained by employing the orientator method. Testicular total volume, the absolute and proportional volumes occupied by the seminiferous tubules and interstitial tissue, as well as the seminiferous tubule length, were estimated using the point-counting system and the unbiased counting frame principle. All the parameters, with the exception of the interstitial tissue's fractional volume, gradually increased along with gestational age, with the maximum increase especially seen in the late fetal stages. The proportional volume of the interstitial tissue, on the other hand, showed a decreasing trend along with increasing gestational age. The absolute volume of the testes, of the seminiferous tubules and of the interstitial tissue, and the length of the seminiferous tubules showed a significant (p< 0.05) positive linear correlation with gestational age. Several similarities were observed with human testicular embryogenesis. The stereological data emerging from the present study might prove useful as basic contribution to the fields of andrology and embryology and stimulate further research in these areas.  相似文献   

7.
旨在克隆奶牛核受体亚家族1 D组成员1(nuclear receptor subfamily 1 group D member 1,NR1D1)基因的蛋白编码序列(coding sequence, CDS),构建其真核表达载体，预测分析NR1D1基因的生物学功能，并检测该基因的组织表达谱及其在卵巢组织的表达分布。本研究以5头健康的24月龄雌性奶牛为试验动物，PCR扩增带有同源臂的奶牛NR1D1基因CDS区全长片段，利用同源重组法将目的片段连接至线性化的pcDNA3.1-Puro-N-3HA空载体，构建重组质粒，利用酶切、PCR及测序对重组质粒进行鉴定。结合质粒测序结果，利用生物信息学软件预测分析该基因的生物学功能。将鉴定正确的质粒转染至HEK293T细胞，利用实时荧光定量PCR(qPCR)与Western blotting技术检测NR1D1基因在mRNA与蛋白水平的表达。此外，利用qPCR检测奶牛NR1D1基因的组织表达谱，并利用免疫组织化学技术检测NR1D1蛋白在奶牛卵巢组织的表达分布。结果，成功获得带有同源臂的奶牛NR1D1基因的CDS区全长片段(1 884 bp),酶切、PCR及...  相似文献   

8.
Abstract

Nonlethal sampling techniques have previously been evaluated for detection of a variety of viral salmonid pathogens. However, many of these studies have used molecular assays in lieu of widely accepted cell culture techniques to evaluate the sampled tissues. Samples were collected from female steelhead Oncorhynchus mykiss broodstock using three potential nonlethal sampling methods (mucus/skin scrape, pectoral fin clip, and gill tissue biopsy) and evaluated for the presence of infectious hematopoietic necrosis virus (IHNV) via cell culture techniques. The results were compared with those from samples collected using a standard lethal sampling method (pooled anterior kidney and spleen tissues) applied to the same fish. Of the three nonlethal sampling techniques that were evaluated, fin clipping was the easiest and least invasive method. Furthermore, fin tissue was as sensitive as or more sensitive than kidney/spleen tissue for detecting IHNV in this population of fish. However, with the exception of gill tissue, the nonlethal samples did not appear to be appropriate surrogates for lethally collected tissues with regard to identifying an active infection in a particular fish. Nevertheless, nonlethal sampling coupled with cell culture appears to be suitable for helping to define the IHNV status of a steelhead population.

Received July 27, 2016; accepted December 11, 2016 Published online March 21, 2017  相似文献   

9.
The aim of this study was to assess cellular proliferation using silver-stained nucleolar organizer regions (AgNOR) and the proliferating cell nuclear antigen (PCNA) in various tissues in the prostate of ram lambs implanted with increasing zeranol doses and to compare the sensitivity of different tissues of lamb prostate to zeranol. Twenty-four Akkaraman lambs were implanted with increasing zeranol doses, including 12 mg (n = 8), 24 mg (n = 8) and 96 mg (n = 8), with eight lambs serving as controls. After 33 days, the prostate tissues of the lambs were stained using AgNOR and PCNA techniques. The prostate tissues were divided into two compartments--the epithelial tissues, including glandular acinus, collecting duct and penile urethra, and the non-epithelial tissues, including interstitial tissue and striated muscle. AgNOR dots and PCNA index on each prostatic tissue were counted under a light microscope and were evaluated statistically. AgNOR staining in the treatment groups showed a higher score in the non-epithelial tissues than the epithelial components, whereas the PCNA index was significant in the epithelial tissues and non-epithelial tissues had very low PCNA immunostaining. According to the PCNA index, collecting duct epithelium showed more sensitivity to increasing zeranol doses and according to AgNOR counts, there was no difference of sensitivity to zeranol among tissues of the same origin. Both AgNOR counts and PCNA indexes seem to be valuable proliferating markers for the epithelial components of ram prostate, but PCNA index had no significance in relation to the non-epithelial components in contrast to AgNOR counts. Therefore, the controversial results arising from the combined use of both techniques as proliferating markers for the ram prostate should be considered in further studies.  相似文献   

10.
The effect of supplementing diets with various levels of vitamin D3 to provide 0, 0.5, 1, and 5 million IU/(steer x d) for 8 d before slaughter on the mineral content and localization of Ca in LM and muscle fragments was studied during the postmortem aging process. Twelve feedlot steers of three biological types were given access to the four levels of vitamin D for 8 d before slaughter. Differential centrifugation techniques were used to determine the concentrations of minerals relative to protein in different muscle fragments on d 3 and 21 postmortem. Electron microscopy visualization of bound Ca indicated that vitamin D3 mobilized Ca from the sarcoplasmic reticulum and transverse tubule system into the myofibrils. Bound Ca was concentrated near the Z-line at the A-band/I-band juncture within the sarcomere. Supplementing steers with 1 and 5 million IU/(steer x d) of vitamin D3 increased (P < 0.05) Ca, P, and Mg concentrations per unit of protein in the cytosol. Soluble cytosolic Ca concentrations were greater (P < 0.05) on d 21 than on d 3 postmortem only when steers were supplemented with 5 million IU/d. Concentrations of Ca, P, and Mg in isolated tissues were increased (P < 0.05) in nuclei and myofibrilar proteins by supplementing steers with 1 and 5 million IU/ (steer x d) of vitamin D3. All supplemental vitamin D3 treatments also increased (P < 0.001) Mg concentrations in the cytosol, regardless of aging treatment, and increased Mg concentrations (P < 0.04) within the mitochondria at d 3 postmortem. Thus, supplementation of feedlot steers with vitamin D3 at levels of 0.5 to 5 million IU/(steer x d) increased Ca concentrations within respiring muscle, resulting in increased bound tissue Ca concentrations. When the respiring muscle was converted to meat, the increased bound tissue Ca resulting from vitamin D3 treatment released Ca concentrations into the cytosol during aging (P < 0.05). Results of this study indicate that vitamin D3 supplementation increased total cytosolic Ca, P, and Mg concentrations in meat.  相似文献   

11.
The aim of this study was to detect the differential expression of miR-383 in yak and cattle-yak testis,and explore the important roles of miR-383 in spermatogenesis.Using the testis tissue of mature yak and cattle-yak as experimental material,Real-time PCR was performed to detect the miR-383 expression.miR-383 targets were obtained using TargetScan and miRanda database,and biological functions of these target genes were predicted by David and Gene Ontology online softwares.The miR-383 had significantly differential expression in F1 testis tissue between yak and cattle-yak(P<0.05),and it had the highest expression level in yak testis tissues.131 target genes were obtained by target gene prediction which involved in cell proliferation,apoptosis,regulation of nucleic acid synthesis and other processes by GO analysis and KEGG pathway analysis.The results showed that miR-383 might participate in the regulation of germ cell differentiation and spermatogenesis,which would provide clues and theoretical basis for studying the function and regulation mechanism of miR-383 in cattle-yak germ cell.  相似文献   

12.
本研究旨在分析miR-383成熟体在雄性牦牛和犏牛F1代睾丸组织中的差异表达,以了解其在精子形成中的重要作用.以发育成熟的牦牛和犏牛睾丸组织为材料,利用实时荧光定量PCR方法检测miR-383成熟体在牦牛及犏牛睾丸组织中的差异表达情况;利用TargetScan和miRanda数据库获得miR-383的靶基因集合,通过David和Gene Ontology在线软件分析miR-383靶基因的生物学功能.实时荧光定量PCR结果显示miR-383成熟体在牦牛和犏牛F1代睾丸组织中表达显著差异(P<0.05),在牦牛睾丸组织中表达量较高,靶基因预测共获得131个靶基因,GO分析及KEGG信号通路分析得到miR-383靶基因功能,这些靶基因参与了细胞增殖、凋亡、核酸合成调控等过程.结合miR-383成熟体的表达差异和靶基因功能预测结果,miR-383可能参与生殖细胞的分化及精子形成调控,为miR-383在犏牛生殖细胞中的功能与调控机制研究提供了理论依据.  相似文献   

13.
以来自内蒙古的野生扁蓿豆种质资源32个居群成熟叶片为试验材料,采用石蜡制片法,应用光学显微技术测定了叶片上表皮厚度、叶片下表皮厚度、叶片厚度、维管束直径、栅栏组织厚度、海绵组织厚度、栅海比、叶片栅栏组织结构紧密度和叶片海绵组织结构疏松度等与抗旱相关的解剖结构特征参数,并进行统计分析和抗旱性综合评价。结果表明,9个指标均表现差异极显著;叶片上表皮厚度、叶片下表皮厚度、叶片厚度、栅栏组织厚度和海绵组织厚度之间呈极显著相关和显著相关关系;第3个主成分的特征值为1.621,累计贡献率达86.67%,已超过85%,前3个主因子基本上能概括9个指标的主要信息;采用UPGMA法将32个居群分成五大类。隶属函数法综合评价抗旱能力显示16和27号居群的抗旱性最强,22号居群抗旱性最差。  相似文献   

14.
Mice experimentally infected with challenge virus standard rabies virus as well as skunks and foxes experimentally infected with street rabies virus were used to demonstrate rabies viral antigen in paraffin-embedded tissue by the peroxidase-antiperoxidase method. Tissues fixed with different fixatives (10% formalin, Bouin's, acetone, ethanol) for various times and fresh frozen tissues were stained by the fluorescent antibody and the peroxidase-antiperoxidase method. Formalin- and Bouin's-fixed tissues were tested with and without use of digestive enzyme (pepsin). The results demonstrated that a procedure using formalin-fixed paraffin-embedded tissue treated with pepsin and stained by peroxidase-antiperoxidase was the best method for both preservation of morphological details and demonstration of antigen.  相似文献   

15.
Monokines are glycoproteins, synthesised by macrophages, which exert various effects on the organism. The most important monokines are interleukin (IL)-1alpha, IL-1beta, tumor necrosis factor (TNF)-alpha and IL-6. This paper reports on immunohistochemical techniques developed for the detection of IL-alpha, IL-1beta, IL-6 and TNF-alpha in fixed and paraffin-embedded pig tissues (spleen, lymph nodes, thymus, liver and kidney). Different fixatives (buffered formalin, acetic formalin, paraformadehyde-lysine-periodate and Bouin solution), and antigen unmasking techniques (permeabilisation with Tween 20, pronase enzymatic digestion and microwave-citrate buffer) were used. We describe different protocols for detection of monokines using polyclonal antibodies against the studied monokines. No signal was obtained with monoclonal antibodies against pig-TNF-alpha and human IL-1alpha. Bouin solution was shown to be the best fixative for immunohistochemical detection of IL-1alpha, TNF-alpha, and IL-6, using permeabilisation with Tween 20 as an unmasking antigen method. Acetic formalin was shown to be the best fixative for IL-1beta detection, not needing antigen retrieval techniques. Macrophages were identified as the main cytokine-producing cells, although other types of cells also stained positively to some cytokines. These techniques represent valuable tools for studies of the pathogenesis of viral and bacterial diseases, and of the immune system of the pigs.  相似文献   

16.
Several phenotypic as well as genotypic methods have been published describing the detection of central nervous system (CNS) tissues that are part of the bovine spongiform encephalopathy (BSE) risk material in food products. However, none of these methods is able to differentiate between CNS tissue of the banned ruminant species and tissues of other animal species. A quantitative and species-specific real-time RT-PCR method has been developed that enables the reliable identification of CNS tissues in meat and meat products. This method is based on a messenger (m)RNA assay that uses bovine, ovine and caprine glial fibrillary acidic protein (GFAP) encoding gene sequences as markers. The in-house validation studies evaluated the tissue specificity of up to 15 bovine tissues and the standardization of absolute as well as relative quantitative measurement. The specific amplification of spinal cord and brain tissue GFAP cDNA has been shown previously. In addition, two commercially available ELISA kits were used for the comparative analysis of artificially contaminated minced meat. Small quantities of bovine brain that had been stored over the recommended period of 14 days were examined. The real-time PCR method proved to be suitable for the detection of 0.1% CNS tissue. No false negative results were observed. The quantitative detection of GFAP mRNA using real-time RT-PCR seems a suitable tool in routine diagnostic testing that assesses the illegal use of CNS tissue in meat and meat products. The stability of the selected target region of the GFAP mRNA also allows the detection of CNS tissues after the meat has been processed.  相似文献   

17.
2007～2008年国际反刍动物营养研究进展 Ⅷ.维生素营养   总被引：1，自引：1，他引：0  
作者综述了2007～2008年脂溶性和水溶性维生素在反刍动物上的研究进展。VA可提高反刍动物的免疫功能,影响牛肉品质和反刍动物脂肪组织的发育,不同来源的VA的生物学活性有差异;VD3可提高奶牛免疫力,VD3的吸收率优于VD2,提高血清中25(OH)D3的浓度,影响牛肉品质;VE可提高产奶量,降低乳房炎、胎衣不下等疾病的发病率,刺激生殖器官发育,改善牛肉品质。烟酸不仅能缓解热应激对奶牛生产性能的影响,还能减少血浆中游离脂肪酸的浓度,从而减少脂肪肝的发生。VB12能促进葡萄糖的代谢,提高初生羔羊的生命力。胆碱能提高奶牛的生产性能,增加产奶量、乳蛋白和乳脂,并影响奶牛的繁殖性能。经过包被的VC有较好的过瘤胃能力,能更多的在肠道被吸收,促进奶牛对VC吸收。  相似文献   

18.
Qi X  Yang X  Cheng A  Wang M  Zhu D  Jia R 《Avian diseases》2008,52(2):338-344
To better understand the pathogenesis of duck virus enteritis (DVE), the levels of viral DNA in various tissues of ducklings during acute stage of virulent duck enteritis virus (DEV) infection were investigated by using quantitative real-time polymerase chain reaction. The results show that the viral levels of DEV in systemic organs have a close correlation with the progression of disease. The rapid dissemination and active replication of virulent DEV in multiple systemic organs at the early phase of acute infection accelerate the progression of disease. The levels of viral DNA increase sharply soon after developed clinical signs of disease, and the extent of increase and the magnitude of DEV DNA load in various tissues of ducklings after the exhibition of clinical signs may be a critical determinant of the outcome of DEV infection. The relatively high levels of DEV in bursa and small intestine tissues of dead ducklings most likely reflect the abundance of target epithelial and lymphoid cells in these tissues, which therefore play a key role in the pathogenesis of acute DVE and manifest as severe tissue lesions on the bursa and small intestine.  相似文献   

19.
Reason for performing study: The relationship between mechanical behaviour and microscopic structure of the laminar junction of equine hooves under testing conditions requires elucidation. Objectives: To determine mechanical parameters and 2D length density of profiles of secondary lamellae of the laminar junction in the dermal region and to assess possible correlations. Methods: Specimens (25 samples in total) of the laminar junction were taken from front, quarter and heel parts from 3 equine hooves and exposed to a uniaxial tensile test until rupture to obtain Young's moduli of elasticity, ultimate stress and strain. Neighbouring specimens to those used for the biomechanical experiment were processed histologically to assess the length density of laminar junction basement membrane using stereological grids. Results: The estimated median (interquartile range) length density of the laminar junction basement membrane was 0.024 (0.020–0.027)/µm. Young's modulus of elasticity was 0.15 (0.11–0.35) MPa in the small deformation region, and 7.58 (6.14–8.68) MPa in the linear region was. The ultimate stress was 1.67 (1.41–2.67) MPa, and the ultimate strain was 0.50 (0.38–0.70). The Young's modulus of elasticity in the region of small deformations has a moderate correlation with the length density of the laminar junction basement membrane. Conclusions: As with most soft biological tissues, the laminar junction has a nonlinear mechanical behaviour. Within the range of small deformations, which correspond to physiological loading of the laminar junction, a higher length density of the laminar junction basement membrane is correlated with a higher resistance of the laminar junction against high stresses transmitted from the distal phalanx to the hoof wall. Potential relevance: The condition of the laminar junction apparatus may be easily quantified as the length density of profiles of secondary dermal lamellae. This quantification provides a simple tool that could be used for comparing the proneness of the various parts of the laminar junction to initial stages of laminitis.  相似文献   

20.
【目的】了解let-7a-5p的生物学信息,初步探索其在静原鸡肌肉组织中的功能作用。【方法】利用生物信息学方法对转录组测序获得的静原鸡let-7a-5p的成熟序列进行保守性分析,使用miRDB、TargetScan和miRanda在线软件预测let-7a-5p的靶基因,取其交集进行GO功能和KEGG通路富集分析,并利用实时荧光定量PCR检测let-7a-5p在静原鸡公鸡和母鸡各组织中的表达水平。【结果】let-7a-5p的成熟序列在各物种间高度保守,共预测得到38个基因集合。在生物过程、细胞组分和分子功能中靶基因富集较多的GO条目有细胞过程、生物调节、细胞、结合和催化活性;靶基因显著富集于聚酮糖单元生物合成、AGE-RAGE信号通路在糖尿病并发症中的作用、泛素介导的蛋白水解、TGF-β信号通路和加压素调节的水重吸收信号通路,且富集最多的是代谢途径、AGE-RAGE信号通路在糖尿病并发症中的作用、泛素介导的蛋白水解和MAPK信号通路。let-7a-5p-靶基因互作主要通过作用于靶mRNA的3'-UTR抑制表达或使其降解。组织表达发现,let-7a-5p在静原鸡公、母鸡的心脏、肝脏、脾脏、肺脏、肾脏、胸肌和腿肌组织中均有表达,且在胸肌和腿肌中表达量均较高。【结论】let-7a-5p可能靶向调节靶基因的表达,从而参与静原鸡肌肉生长发育,为进一步探究let-7a-5p的功能和作用机制提供理论依据。  相似文献   

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