D-半乳糖促小鼠衰老模型的衰老程度与自然生长鼠龄的比较

为探讨不同剂量D-半乳糖对小鼠的促衰程度及其与自然生长鼠年龄的对应关系,应用改进的邻苯三酚自氧化法对自然生长鼠和D-半乳糖促衰小鼠肝脏中SOD活力进行了测定、比较和分析。结果表明,自然生长鼠约从11月龄开始进入明显的衰老期,至17月龄衰老程度较高,至21月龄时小鼠的衰老程度极高。还总结了不同剂量D-gal促衰鼠与自然生长鼠年龄的对应关系。     

OSTEOCHONDROSIS IN THE LATERAL FEMORAL CONDYLES OF A HORSE

Osteochondrosis of the lateral femoral condyles was diagnosed radiographically in an 8-month-old, female Arabian horse, which had been presented with a hindlimb lameness. The diagnosis was confirmed by gross and microscopic pathology. The location of the lesions was considered unusual for osteochondrosis in the horse.     

堆型艾美球虫3-1E基因在大肠杆菌中的可溶性表达及鉴定

将堆型艾美球虫（E．acervulina）3-1E基因克隆至pET-32a（＋）载体中。转化大肠杆菌BL21，在37℃下，用终浓度为1.0mmol／L的IPTG诱导表达，得到相对分子质量约为38500的重组蛋白。Western blot检测证实，该重组蛋白可以与特异性抗血清结合。在20℃条件下，以终浓度分别为2．0、1．0、0．5mmol／L的IPTG进行诱导．当IPTG终浓度为0．5mmol／L时，以可溶性形式存在的目的蛋白含量最高。     

Biomechanical Comparison of Two Alternative Tibial Plateau Leveling Osteotomy Plates with the Original Standard in an Axially Loaded Gap Model: An In Vitro Study

Objective— To compare the axial compression stiffness of osteotomized canine tibiae stabilized with Slocum, Securos, or Synthes plates after a tibial plateau leveling osteotomy (TPLO) procedure. Study Design— In vitro, paired comparison of cadaveric tibial constructs subjected to mechanical testing under an axial load. Sample Population— Canine tibiae (n=16 pairs) from skeletally mature male and female dogs of various breeds (18–55 kg). Methods— Tibial pairs (n=16) were randomly assigned to 1 of 2 study cohorts (n=8 pairs/cohort): cohort 1, tibial osteotomy stabilization with a Slocum or a Securos plate, or cohort 2, tibial osteotomy stabilization with a Slocum or a Synthes plate. One tibia from each pair was stabilized with 1 of each plate design assigned to the cohort after TPLO. A 3.2 mm osteotomy gap was maintained during plate application in all constructs. Load and axial displacement were recorded while constructs were loaded to 2000 N in axial compression. Failure loads were not reported because no distinct yield point or failure point was evident within the load range for many specimens. Failure modes were recorded for each construct, and photographs of typical failures were obtained. Stiffness (N/mm) was calculated from load–displacement curves. Paired comparisons of mean stiffness were performed within study groups using a paired t‐test. Significance was set at P<.05. Results— The mean construct stiffnesses for the Slocum (383±183 N/mm) and Securos (258±64.1 N/mm) constructs were not significantly different (P=.164; power=0.566). The mean construct stiffness for the Synthes constructs (486±91.0 N/mm) was significantly greater than that of the Slocum constructs (400±117 N/mm); P=.0468. Modes of failure for the Slocum (16/16) and Securos (8/8) constructs included plastic deformation of the implant with valgus deformity combined with fibular luxation (2/16 Slocum; 1/8 Securos) or fibular fracture (2/16 Slocum; 4/8 Securos). Most Synthes constructs underwent elastic deformation (7/8). One Synthes construct fractured in the saggital plane through the tibial plateau depression at the point of load application. Conclusions— The Slocum and Securos plate/tibia construct have similar stiffness, whereas the Synthes/tibia constructs are significantly stiffer than the Slocum/tibia constructs. Modes of fixation failure observed in this model were consistent with TPLO fixation failures observed clinically. Clinical relevance— Construct stiffness in axial load varies with implant type. Implants that confer higher stiffness to the construct may result in greater fixation stability in tibial metaphyseal osteotomies.     

药用植物紫花地丁的分子鉴定及其亲缘关系研究

运用RAPD技术对传统药用植物紫花地丁（Viola yedoensis）及其7个近缘种进行了分析,筛选出的6个随机引物在8个种39份材料中共扩增出163个位点,其中161个位点具有多态性,占98.7%;在扩增产物中有7条特异性扩增条带可用于紫花地丁的鉴定。根据RAPD结果计算的各种间遗传距离的范围为0.022 7-0.424 4,斑叶堇菜与细距堇菜的亲缘关系最近,它们间的遗传距离最小（0.022 7）,而与紫花地丁遗传距离最近的是早开堇菜,为0.147 1。在所研究的类群中,维西堇菜与裂叶堇菜的遗传距离最大（0.424 4）。根据遗传距离所作的聚类分析结果与紫花地丁等8种植物的形态分化相一致,此外,同一种的不同个体总是先聚在一起,反映了种内不同个体在遗传组成上相对一致性。研究结果表明,RAPD在种内是相对稳定的,能快速、准确地应用于紫花地丁的分子鉴定及其与近缘类群关系的研究。     

鸡新城疫病毒河北分离株F基因的分子特性分析

通过毒力测定、RT-PCR及F基因的序列测定与遗传进化分析,对2005-2008年从河北省部分地区的发病鸡群中分离到的10株新城疫病毒（NDV）进行了研究。各分离株经典毒力测定结果显示：MDT在37.6～54.4h之间,ICPI在1.71～2.0之间,IVPI值在2.16～2.8之间,均为新城疫病毒强毒株特征。F基因的序列测定表明,分离株之间的核苷酸序列具有77.4%～98.0%的同源性,与疫苗株Lasota的同源性为87.0%～98.9%,与国内标准强毒株F48E9同源性为89.7%～98.9%。推导其氨基酸序列分析表明,8个分离株的F蛋白的裂解位点氨基酸组成为112 R-R-Q-K-R-F117,具有NDV强毒株特征,与毒力测定结果相符,2个分离株的F蛋白的裂解位点氨基酸组成为112 G-R-Q-G-R-L117,与弱毒株特征相符。F基因分型和同源性比较显示：目前河北新城疫的流行以基因Ⅶ型为主（占70%）,同时兼有基因Ⅱ型（占20%）和基因Ⅸ型（占10%）。     

湛江地区肉牛棚舍环境评价

为了解热带地区双坡式棚舍散栏饲养模式下的环境特征,在湛江测定了肉牛舍的温度、湿度、气流、PM10等指标。结果表明:棚舍内外温湿度几乎相同,换气量超过了最大换气量,PM10优于卫生标准,温湿指数范围为78～85,处于热应激状态。考虑了气流影响的体感温度范围为25～30.5℃,晴天的14:00和19:00,风速为0.95～1.30 m/s,气流显著地减少了高温的影响。棚顶隔热能力差,晴天棚顶内外表面温差不足1℃,棚顶内表面最高可达60.8℃,此时的舍内气温仅为33.4℃,棚舍自然换气调节是环境控制的主体。     

猪伪狂犬病病毒gE抗体胶体金免疫层析检测方法的建立及应用

以纯化的抗人红细胞单链抗体(ScFv)-猪伪狂犬病病毒(PRV)gE蛋白双功能融合蛋白为诊断抗原和胶体金标记物,以羊抗猪IgG包被硝酸纤维膜作为质控带,制作检测猪伪狂犬病毒gE抗体的双抗原胶体金试纸条。利用方阵滴定试验筛选出金标抗原最佳工作浓度为17.6μg,检测线诊断抗原最佳标记量为1.76μg,血清最佳稀释度为1∶10,作用时间15 min,与猪瘟病毒(CSFV)、猪细小病毒(PPV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪乙型脑炎病毒(JEV)、猪布鲁菌(Brucella)阳性血清和PRVgE缺失疫苗接种的猪免疫血清检测线均不出现红色条带,与PRV标准阳性血清反应检测线出现红色条带。试纸条操作简单,肉眼于15 min内可判定结果;试纸条在室温保存6个月,其特异性和敏感性没有明显变化;与美国IDEXX和法国LSI gE-ELISA抗体检测诊断试剂盒检测结果比较,1 164份猪血清的符合率均为90.55%。制备的胶体金试纸条具有操作简便、敏感性和特异性较高的特点,可用于PRV野毒感染的快速筛查。     

Oral vaccination of brushtail possums (Trichosurus vulpecula) with BCG: immune responses,persistence of BCG in lymphoid organs and excretion in faeces

AIMS: To determine immune responses, and the localisation and persistence of Mycobacterium bovis bacille Calmette-Guérin (BCG) in gut-associated lymphoid tissues (GALT) and other organs in possums vaccinated orally with lipid-formulated BCG vaccine. To determine the duration of excretion and longevity of survival of BCG in the faeces of vaccinated animals.

METHODS: Possums (n=28) were vaccinated with lipid-formulated BCG (1 x 10 ⁸ colony forming units (cfu) of formulated BCG) by the oral route. Control possums (n=17) were fed oral bait pellets containing formulation medium only. Possums were sacrificed at 3 days and at 1, 3, 6 and 8 weeks after vaccination or ingestion of bait. Proliferation responses to bovine purified protein derivative (PPD) were measured in lymphocytes from blood and mesenteric lymph nodes (MLN) and samples of lung, spleen, liver, MLN and Peyer's patches (PP) were cultured for the presence of BCG. The number of BCG organisms excreted in faeces and the duration of excretion were determined in eight vaccinated possums and eight control possums over a 3-week period. In a separate experiment, a further six possums were vaccinated with oral BCG vaccine (5–10 x 10 ⁸ cfu BCG/possum) and their faeces collected over 48–72 h, for culture of BCG. The longevity of survival of BCG in these faeces was determined by storing faecal samples (n=12) under three different conditions: in an incubator (22.5°C), and conditions which simulated the forest floor and open pasture. A proportion (1–2 g) of these faecal samples was collected after storage for 1, 3, 5, 8 or 20 weeks, and cultured for BCG.

RESULTS: Possums vaccinated orally with BCG vaccine showed strong proliferation responses to bovine PPD in peripheral blood lymphocytes at 6–8 weeks post-vaccination (p.v.). Positive lymphocyte proliferation assay (LPA) responses to bovine PPD were first evident in MLN at 3 weeks p.v. BCG was cultured from MLN and PP in a proportion of animals at 3–8 weeks p.v. BCG was not cultured from sections of spleen, lung or liver at any time p.v. BCG was recovered in low to moderate numbers from the faeces of vaccinated possums for up to 7 days, and maximal numbers were cultured in faeces collected 48–72 h p.v. After storage for 1 week, BCG was cultured from all faecal samples placed in the incubator and from a proportion of faeces exposed to conditions similar to those on the forest floor and pasture. With the exception of one faecal sample stored under forest floor conditions which was culture-positive for BCG at 3 and 5 weeks, BCG was not cultured from any other faecal sample stored for more than 1 week.

CONCLUSIONS: Ingestion of oral BCG vaccine by possums was associated with the development of strong cell-mediated immunity in both blood and MLN. Following oral vaccination with BCG, the organisms were localised and persisted in GALT but did not spread to the spleen, liver or lungs. BCG was shed in low to moderate numbers in the faeces for up to 7 days p.v. The viability of BCG excreted in faeces decreased rapidly, particularly when faeces were exposed to an open pasture environment. Oral vaccination of possums with formulated BCG is unlikely to result in undue contamination of the environment with BCG.