8种中药成分体外抗犬细小病毒的研究

本研究旨在观察8种中药成分黄芪多糖（APS）,白术多糖（RAPS）,黄芩多糖（BSPS）,金银花多糖（FLPS）,栀子苷（CS）,苦参生物碱（SFA）,黄芪皂苷（AS）和金银花黄酮（FLF）体外对犬细小病毒（CPV）感染细胞能力的影响。分别将8种中药成分通过先加中药再接病毒、病毒和中药感作后加入和先接种病毒再加中药3种加药方式加入到培养24h的F81细胞中,于病毒接种后72h用MTT法测定F81细胞活性以评价各中药成分对F81细胞抵抗犬细小病毒感染的影响。结果,在第1种加药方式下,APS、RAPS、BSPS、FLPS抗CPV作用较强;在第2种加药方式下CS的抗病毒作用最为显著;在第3种加药方式下CS、SFA抗病毒作用最为显著。结果表明,8种中药成分均能不同程度地促进F81细胞抵抗病毒感染,部分中药成分的抗病毒作用与加药方式有关且呈一定的量效关系。     

犬瘟热患犬中性粒细胞cAMP PDE活性变化及中药作用研究

研究犬瘟热患犬中性粒细胞(PMN)中环磷酸腺苷-磷酸二酯酶（cAMP-PDE）的活性变化及中药对其活性的体外调节作用，为抗炎药研究和病毒性炎症的中药治疗积累资料。提取犬瘟热患犬（n=9）和正常犬(n=6)的中性粒细胞制备cAMP-PDE样品，通过高效液相色谱(HPLC)法测定其活性及中药对其活性的影响。结果显示，犬瘟热患犬组中性粒细〖JP2〗胞cAMP-PDE活性（18.55%±2.66%）显著（P<0.05）高于正常组（11.54%±5.16%）。解表药等5类114味中药中芦根（77.27%）、胡黄连（70.41%）、女贞子（65.50%）、香薷（65.16%）、贝母（61.43%）、黄芩（60.67%）、淫羊藿（60.34%）、黄芪（59.13%）、青蒿（56.93%）、秦皮（55.55%）、辛夷（55.03%）、甘草（54.67%）、牛蒡子（53.40%）、麻黄（53.32%）14味对cAMP-PDE活性具有明显抑制作用。结果表明，中性粒细胞cAMP-PDE活性在犬瘟热患犬炎症过程中具有重要作用，对cAMP-PDE活性有明显抑制作用的中药对其炎症可能具有疗效，同时，中药抑制中性粒细胞中cAMP-PDE活性可能是其抗炎作用机理之一。     

中药成分对培养的鸡脾脏淋巴细胞增殖的影响

为筛选增强免疫作用较好的中药成分和研制新型复方中药成分免疫增强剂，比较了4种浓度的当归多糖（CAPS）、黄芪多糖（APS）、板蓝根多糖（IRPS）、淫羊藿多糖（EPS）、蜂胶多糖（PPS）、淫羊藿黄酮（EF）、蜂胶黄酮（PF）、黄芪皂苷（AS）和人参皂苷（GS）9种中药成分对培养的鸡脾脏淋巴细胞增殖的影响。结果表明，多数中药成分能促进淋巴细胞增殖，其中APS、EPS、GS和AS等在一定浓度既能单独又能协同ConA或LPS刺激淋巴细胞增殖，以EPS和APS作用最强。同时根据浓度试验结果，选出各药的合适浓度，进一步比较它们作用的差异，发现EPS和GS效果最好。这些中药成分可以作为复方中药成分免疫增强剂的组分药。     

黄苠多糖对细小病毒感染犬血常规与脂质过氧化的影响及疗效

将临床收集确诊的40只细小病毒病犬分为黄芪多糖组和常规治疗组，对黄芪多糖是否具有治疗犬细小病毒病的作用进行试验研究。并以犬血常规变化来检验其疗效。结果表明：细小病毒病以2～5月龄犬发病较多，并以肠炎型为主。犬感染细小病毒后白细胞数、红细胞数、血红蛋白含量、红细胞压积均低于正常值范围。黄芪多糖能够升高病犬白细胞数、红细胞数、血红蛋白含量，除红细胞数外，在治疗第6天均恢复到正常水平。黄芪多糖能够升高病犬血清中SOD的活性，治疗第6天与基础值比较差异极显著（P〈0．01）；同时能够降低病犬血清中MDA的含量，在治疗第6天与基础值比较差异极显著（〈O．01），与健康对照组相近。黄芪多糖组治愈率为90％．常规治疗组治愈率为70％。     

黄芪甲苷对染镉鸡脾淋巴细胞膜氧化损伤的保护作用

探讨黄芪甲苷对镉致鸡脾淋巴细胞膜氧化损伤的保护效果.设对照组、0.5%二甲基亚砜(DMSO)组、镉(10μmol/L)组和镉(10μmol/L)加黄芪甲苷(分别为5、10和20 mg/L)组,进行体外培养淋巴细胞,分别培养24、48和72 h时收集细胞,提取脾淋巴细胞膜,采用试剂盒比色法,分别检测细胞膜谷胱甘肽过氧化物酶(GSH -Px)和超氧化物歧化酶(SOD)的活性及膜丙二醛(MDA)的含量.DMSO组与对照组相比无显著性差异;镉能明显降低细胞膜GSH-Px和SOD的活性,使MDA含量增加;添加黄芪甲苷的各组中细胞膜GSH-Px和SOD的活性明显升高,MDA含量降低,其影响较为显著的是10 mg/L的黄芪甲苷质量浓度组.结果表明:黄芪甲苷可以保护染镉后鸡脾淋巴细胞膜的氧化损伤作用.     

板蓝根、黄芪等中药活性提取物对猪繁殖与呼吸综合征病毒体外作用的研究

本试验利用Marc-145细胞体外培养系统，通过观察细胞病变效应（CPE）来评价板蓝根、黄芪等中药活性提取物成分体外抑制猪繁殖与呼吸综合征病毒（PRRSV）对细胞的感染作用，并通过改变加药方式（先加药物后接种病毒、先接种病毒后加药物、病毒和药物感作后同时加入），初步探讨中药活性提取物的抗病毒机制。结果表明。在安全浓度范围内，板蓝根水提物体外对PRRSV具有显著的直接杀灭作用；连翘、黄芪水提物和黄芪多糖体外对PRRSV均具有明显的阻断和抑制作用，为筛选抗PRRSV中药制剂提供了理论依据。     

3种中药活性成分对猪圆环病毒2型疫苗免疫效果的影响

为筛选对猪圆环病毒2型(PCV2)疫苗有显著免疫增强作用的中药活性成分,本研究将66头14日龄猪随机分为11组,分别为免疫对照组、空白对照组和9个给药组。饲养至21 d时,除空白对照组外,其余各组注射2 mL PCV2灭活疫苗。在免疫前3 d和免疫后3 d,9个给药组分别灌喂3个剂量水平(高剂量12 mg,中剂量8 mg,低剂量4 mg)野黄芩苷、苦参碱和黄芪多糖溶液5 mL,免疫对照组和空白对照组灌喂等体积生理盐水。分别在免疫后第7 d、14 d、21 d和28 d,各组随机抽取5头猪静脉采血,测定PCV2抗体水平、特异性免疫球蛋白IgG含量、外周血CD~(3+)含量和CD4~+/CD8~+值、血清IL-2及IFN-γ含量变化。结果表明,3种中药活性成分对PCV2疫苗的免疫效果均有不同程度的增强作用,其中野黄芩苷中剂量组效果最佳,可以作为PCV2疫苗免疫增强剂的候选药。     

黄芪多糖、灵芝多糖益生菌组合对幼犬生长性能与免疫力的影响

本试验旨在验证两种植物多糖-黄芪多糖、灵芝多糖和益生菌组合对幼犬生长性能及免疫力的影响。选取9只3月龄雄性幼犬，配对分配到3个小组，每组每个品种各1只。试验A组幼犬饲喂全价基础幼犬粮+某品牌犬用益生菌+黄芪多糖，试验B组幼犬饲喂全价基础幼犬粮+某品牌犬用益生菌+灵芝多糖，试验C组为对照组，饲喂全价基础幼犬粮+某品牌犬用益生菌，预饲期11d，正式测试期63d。结果显示：两种多糖益生菌组合均未对犬只生长发育造成不良影响，灵芝多糖组合体重增长速度略优于黄芪多糖组合、对照组（P> 0.05）；胃肠消化方面，黄芪多糖组合略优于灵芝多糖组合（P> 0.05），但明显优于对照组，差异显著（P <0.05）；免疫力方面，两种多糖益生菌组合相比，对照组血清IgG抗体、粪便IgA抗体水平差异均不显著（P> 0.05），且两种多糖益生菌组合之间差异不显著（P> 0.05）。     

几种中药成分的免疫增强活性及其作用效果

测定了黄芪多糖等9种中药成分对正常小鼠脾和外周血淋巴细胞增殖反应的影响，并观察了这些中药成分对兔瘟组织灭活疫苗的增强免疫效果。结果表明，人参皂甙、黄芪多糖、淫羊藿多糖、当归多糖和蜂胶黄酮能显著增强伴刀豆球蛋白(ConA)和脂多糖(LPS)诱导的小鼠淋巴细胞增殖，提高免疫兔抗体水平和延长抗体持续时间；板蓝根多糖能增强ConA和LPS的诱导活性，蜂胶多糖能促进ConA的诱导活性，黄芪皂甙和淫羊藿黄酮能促进LPS诱导的淋巴细胞增殖，但它们都不能提高兔瘟组织灭活疫苗的免疫抗体水平。     

铜缺乏对奶牛红细胞及组织细胞膜ATP酶活性的影响

为探讨铜缺乏对奶牛红细胞及组织细胞膜ATP活性的影响，在调查铜缺乏奶牛水土食物链微量元素状态的基础上，首次测度了病区6头发病犊奶牛（Ⅱ组）及该地区20头同年生临床健康奶牛（IH组，血清铜低于0.56mg/kg）红细胞膜Na^ -K^ -ATP酶、Ca^2 -ATP酶及Mg^2 -ATP酶的活性，对20头IH组奶牛进行为期80d的补铜试验（含铜40mg/kg干饲料），以20头健区同年生健康奶犊牛作为对照（HH组，全血铜大于０.85mg/L）。同时测试了剖杀后６头病牛及健康牛组织细胞膜ＡＴＰ酶的活性。结果表明：Ⅱ组奶牛红细胞膜ＡＴＰ酶活性显著低于ＨＨ组及ＩＨ组。补铜后４０d，ＩＨ组奶牛红细胞膜Na^ -K^ -ATP酶均显著升高，病牛肝、脾、肾、大脑、小脑、淋巴结等组织细胞膜Na^ -K^ -ATP酶、Ca^2 -ATP酶活性显著低于对照组。结论：铜缺乏较为严重地影响了奶牛红细胞及组织膜ＡＴＰ酶的活性，抑制了这些组织的正常生理功能。     

Cellular immunophenotyping of exfoliative dermatitis in canine leishmaniosis (Leishmania infantum)

Lymphocyte subsets, major histocompatibility complex (MHC)-II expressing cells and number of amastigotes in the epidermis and dermis were investigated immunohistochemically in 48 dogs with patent leishmaniosis, with or without exfoliative dermatitis (ED) to study the immunopathogenesis of this common cutaneous form of the disease. Skin biopsies were obtained and compared for ED sites (group A, n = 26), normal-appearing skin from the same animals (group B, n = 24), and leishmanial dogs not exhibiting ED (group C, n = 22), and normal controls (group D, n = 22). The CD3+, CD45RA+, CD4+, CD8+ (CD8a+), CD21+, and MHC-II+ cells and leishmania amastigotes were identified immunohistochemically and counted with the aid of an image analysis system. Pyogranulomatous to granulomatous dermatitis, expressed in various histopathological patterns, was noticed in all groups A and B and in half of group C dogs. In the epidermis, the low number of T-cells and their subsets did not differ significantly between groups A and B, but CD8+ outnumbered CD4+ lymphocytes in both groups. MHC-II+ expression on epidermal keratinocytes was intense in the skin with and without lesions from dogs with ED but not in group C dogs. CD3+, CD8+ and MHC-II+ cells were fewer in group C compared to group A and B dogs. In the dermis, CD3+ cells in group A animals were mainly represented by the CD8+. CD45RA+ and CD21+ cells were also seen in high numbers. MHC-II expression, potentially in lymphocytes, fibroblasts, dendritic cells, and macrophages was intense. The numbers of all cellular subpopulations in the dermis were significantly different between the groups, being highest in group A and lowest in group D. In sebaceous adenitis sites, CD4+ outnumbered CD8+ cells in contrast to the neighbouring dermis and the epidermis. The number of CD21+ and CD45RA+ cells was much lower in the inflamed sebaceous glands compared to the dermis. Finally, the number of amastigotes in the normal-appearing skin was significantly higher in the ED dogs (group B) than in those not exhibiting this cutaneous form of the disease (group C).     

PDGFR-β在不同年龄牦牛肺中分布和表达的研究

旨在研究血小板源性生长因子受体-β（PDGFR-β）在牦牛肺中的分布和表达,探讨牦牛肺对高原低氧环境的适应机制。采用免疫组织化学、Western-blot技术和qRT-PCR检测方法对新生、1岁、3岁及6岁牦牛肺中PDGFR-β的准确分布位置及表达量进行研究。结果显示：PDGFR-β主要分布在肺内支气管及其分支的上皮细胞、肺动脉血管内皮细胞及管壁平滑肌细胞中,其表达较强;少量分布在支气管管壁平滑肌细胞与肺泡隔细胞中,其表达较弱。在不同年龄段牦牛肺中PDGFR-β蛋白和mRNA也有不同程度的表达。在蛋白水平上,新生组极显著高于其他3组（P<0.01）;1岁组、3岁组、6岁组两两相比较,差异性不显著（P>0.05）。在mRNA表达水平上,以新生组的表达最强,与其他3组相比较,差异性极显著（P<0.01）;1岁组表达量极显著高于3岁组、6岁组（P<0.01）;6岁组显著高于3岁组（P<0.05）。试验表明,PDGFR-β在不同年龄牦牛气道和肺动脉发育及适应性结构的形成过程中均发挥作用,以新生到1岁龄之间的表现最为明显。     

Scanning electron microscopy of vascular corrosion casts and histologic examination of pulmonary microvasculature in dogs with dirofilariosis

OBJECTIVE: To characterize structural changes in pulmonary vessels of dogs with dirofilariosis. ANIMALS: 8 dogs with dirofilariosis and 2 unaffected control dogs. PROCEDURE: Pulmonary artery pressure was measured in affected dogs, and dogs then were euthanatized. Scanning electron microscopy was used to examine vascular corrosion casts of pulmonary vasculature. Tissue sections of pulmonary vasculature were evaluated by use of histologic examination. RESULTS: Pulmonary artery pressure was higher in dogs with severely affected pulmonary vessels. In tissue sections, dilatation, as well as lesions in the tunica intima and proliferative lesions resulting in constriction or obstruction, were frequently observed in branches of the pulmonary artery. Numerous dilated bronchial arteries were observed around affected pulmonary arteries. Hyperplastic venous sphincters were observed in small pulmonary veins and venules. In corrosion casts, affected pulmonary lobar arteries had dilatation, pruning, abnormal tapering, constriction, and obstruction. In small arteries and arterioles, surface structures representing aneurisms and edema were seen. Bronchial arteries were well developed and extremely dilated, and they formed numerous anastomoses with pulmonary arteries at all levels, from the pulmonary trunk to peripheral vessels. Capillaries in the lungs were dilated with little structural change. Small pulmonary veins and venules had irregular annular constrictions that were caused by hyperplastic smooth muscle cells of venous sphincters. CONCLUSIONS AND CLINICAL RELEVANCE: Scanning electron microscopy of microvascular casts delineated links between the bronchial and pulmonary circulations in dogs with dirofilariosis. Results of scanning electron microscopy provided a structural explanation for the development of pulmonary circulatory disturbances and pulmonary hypertension in dogs affected by dirofilariosis.     

Ultrastructural morphologic evaluation of the phenotype of valvular interstitial cells in dogs with myxomatous degeneration of the mitral valve

OBJECTIVE: To evaluate morphologic changes in valvular interstitial cells of dogs and to find evidence for disease-associated phenotypic changes in these cells. ANIMALS: 5 clinically normal dogs and 5 dogs with severe mitral valve endocardiosis. PROCEDURE: Mitral valve leaflets were evaluated by use of transmission electron microscopy. Differences in cell type and cell location were identified. RESULTS: A change in cell type toward a myofibroblast or smooth muscle cell phenotype was detected, with the smooth muscle cell type being most common. These cells had long amorphous cytoplasmic extensions, fibrillar cytoplasm, incomplete basal lamina, few mitochondria, and eccentrically placed nuclei but lacked smooth endoplasmic reticulum or Golgi complexes. Remaining valvular interstitial cells had heterochromatic nuclei and produced only minimal quantities of collagen. Compared with normal valves, myxomatous valves ha many interstitial-like cells located adjacent to the endothelium. Deeper within the abnormal valves, cells with a heterogenous phenotype formed groupings that appeared to be anchored to adjacent collagen. CONCLUSIONS AND CLINICAL RELEVANCE: Myxomatous degeneration of the mitral valve in dogs is associated with phenotypic alteration, changing from an interstitial to a mixed myofibroblast or smooth muscle cell phenotype. A closer association between interstitial cells and the endothelium is evident in diseased valves. In response to the disease process, valvular interstitial cells of dogs appear to change toward a smooth muscle phenotype, possibly in an attempt to maintain valve tone and mechanical function.     

Increased splenic capacity in response to transdermal application of nitroglycerine in the dog

This study was conducted to determine if application of transdermal 2% nitroglycerine ointment (TDNG) to dogs anesthetized with alpha chloralose would produce splenic dilatation by relaxation of venous smooth muscle. Sonomicrometer crystals were applied to the spleen in each of 15 dogs, and a pressure-measuring catheter was inserted into a splenic vein. The sonomicrometer crystals permitted measurement of splenic dimension, which is known to correlate with splenic volume. Ten dogs were given 2.5 cm TDNG/10 kg, and 5 dogs (vehicle controls) were given only petrolatum, both applied to the inner surface of the auricular pinna. Splenic dimension in all dogs receiving TDNG increased significantly (P < .05) by 7.0 +/- 4.8%, whereas splenic dimension in dogs receiving petrolatum did not increase. Splenic venous pressures did not change significantly in either group. Spleens began to dilate 482 +/- 652 seconds after application of TDNG and achieved maximal dilatation at 861 +/- 632 seconds. Splenic dilatation occurred in the absence of elevation of splenic venous pressure, indicating that the dilatation probably resulted from relaxation of splenic smooth muscle. TDNG was absorbed transdermally and produced splenic dilatation in health dogs anesthetized with alpha chloralose. If the spleen is a sentinel for peripheral veins, then TDNG may increase venous capacity, retaining blood from the lungs and thereby functioning to reduce pulmonary congestion and edema in dogs with left-sided heart failure.     

Ultrastruct of the juxtaglomerular apparatus in the dog in a sodium-balanced state.

The ultrastructure of juxtaglomerular apparatuses (JGA) from four dogs in a state of sodium balance were examined. Plasma renin activity and urinary sodium excretion in the dogs were within normal values. At the vascular pole of the glomerulus, circumferentially oriented smooth muscle cells in the wall of the glomerular arteriole were abruptly replaced by modified smooth muscle cells with complicated interwoven cell processes. These modified cells were indistinguishable from the juxtaglomerular cells of the lacis region (extraglomerular mesangium). The basal lamina surrounding these modified smooth muscle cells in the wall of the glomerular arteriole was continuous with the basal lamina of the juxtaglomerular cells of the lacis (extraglomerular mesangium) and the macula densa. The most striking differences in the JGA of the dog, when compared with the JGA of other laboratory animals, are related to the observation that cytoplasmic secretory granules are less abundant in the dog, are generally smaller than the practical resolving capability of the light microscope, and are restricted to cells of the lacis area in the JGA.     

Phenotypical characterization of T and B cell areas in lymphoid tissues of dogs with spontaneous distemper

CD3, CD4, CD5, and CD8 antigen expression of T cells and IgG expression of B cells and canine distemper virus (CDV) antigen distribution were immunohistochemically examined in lymphoid tissues (lymph node, spleen, thymus, and tonsil) of control dogs and animals with spontaneous canine distemper. In addition, CNS tissue of all animals was studied for neuropathological changes and CDV antigen distribution. Based on the degree of depletion distemper dogs were classified into two groups. Group I represented animals with moderate to marked lymphoid depletion, while group II dogs displayed mild or no depletion. CDV antigen was mainly found in lymphocytes and macrophages of group I dogs, whereas CDV expression was most prominent in dendritic cells of group II animals. In group I dogs, a marked loss of CD3, CD4, CD5, CD8, and IgG expression was noticed, hereby loss of CD4+ cells was more prominent than depletion of CD8+ cells. In the lymphoid tissues of group II animals, a significant increase in the number of T and B cells was observed compared to group I dogs. The number of CD3+, CD4+, and CD8+ cells in group II dogs was similar to the findings in controls, however, CD5 and IgG expression was mildly reduced in T and B cell areas, respectively. Additionally, in groups I and II dogs, CD3+ and CD5- T cells were detected in T cell areas. Whether this cell population represents a cell type with autoimmune reactive potential remains to be determined. Surprisingly in group II animals, viral antigen was found predominantly in dendritic cells indicating a change in the cell tropism of CDV during chronic infection and a possible mechanism of viral persistence. The two patterns of lymphoid depletions correlated to two different types of canine distemper encephalitis (CDE). Group I dogs displayed acute non-inflammatory CDE, whereas group II dogs suffered from chronic inflammatory demyelinating CDE, indicating a pathogenic relationship between lymphocytic depletion and inflammatory brain lesions in distemper.     

Immunohistochemical determination of the expression of endothelin receptors in bronchial smooth muscle and epithelium of healthy horses and horses affected by summer pasture-associated obstructive pulmonary disease

OBJECTIVE: To immunohistochemically determine the expression of endothelin (ET) receptors in bronchial smooth muscle and epithelium of healthy horses and horses affected by summer pasture-associated obstructive pulmonary disease (SPAOPD). SAMPLE POPULATION: Tissue specimens obtained from 8 healthy and 8 SPAOPD-affected horses. PROCEDURE: Horses were examined and assigned to healthy and SPAOPD groups. Horses were then euthanatized, and tissue specimens containing bronchi of approximately 4 to 8 mm in diameter were immediately collected from all lung lobes, fixed in zinc-formalin solution for 12 hours, and embedded in paraffin. Polyclonal primary antibodies against ET-A or ET-B receptors at a dilution of 1:200 and biotinylated IgG secondary antibodies were applied to tissue sections, followed by the addition of an avidin-biotin immunoperoxidase complex. Photographs of the stained slides were digitally recorded and analyzed by use of image analysis software to determine the intensity of staining. Two-way ANOVA was used for statistical analysis. RESULTS: The left diaphragmatic lung lobe of SPAOPD-affected horses had a significantly greater area of bronchial smooth muscle that immunostained for ET-A, compared with that for healthy horses. All lung lobes of SPAOPD-affected horses, except for the right diaphragmatic lobe, had significantly greater staining for ET-B receptors in bronchial smooth muscle, compared with results for healthy horses. CONCLUSIONS AND CLINICAL RELEVANCE: This study revealed overexpression of ET-A and, in particular, ETB receptors in the bronchial smooth muscle of SPAOPD-affected horses, which suggested upregulation of these receptors. These findings improve our understanding of the role of ET-1 in the pathogenesis of SPAOPD.