ZBED6基因对巴马香猪脾发育的作用机制分析

ZBED6基因作为一个转录因子,能够与IGF2结合进而调节肌肉的生长发育。但其在脾生长发育中的作用尚不清楚,本研究利用RNA-seq测序技术比较ZBED6基因敲除巴马香猪（ZBED6-KO）和同日龄野生型巴马香猪（WT）的脾组织转录组,探究ZBED6基因对巴马香猪脾组织的发育影响。利用t检验对ZBED6-KO猪和WT猪脾组织大小的表型差异及ZBED6直接调控的靶基因IGF2的表达量进行显著性分析。提取ZBED6-KO猪和WT猪脾组织的总RNA,在Illumina Hiseq 2500平台进行RNA-seq分析。以猪Sus scrofa11.1为参考基因组,用转录组分析的标准流程筛选ZBED6-KO猪和WT猪脾组织中的差异表达基因。用DAVID在线网站对差异表达基因进行GO和KEGG富集分析。然后,随机选取7个差异表达的基因,利用实时荧光定量PCR技术验证测序结果的准确性与可靠性。结果显示,与WT猪相比,ZBED6-KO猪脾的重量和IGF2基因表达量均显著增加（P<0.05）,表明ZBED6基因的敲除对巴马香猪脾组织的生长发育有一定的促进作用。测序结果显示,各样本至少获得4G的数据量,每个样本的Clean Ratio及Q30比率均在90%以上,83.94%以上的reads可比对在猪的参考基因组上,表明测序数据质量良好,真实可靠;对测序数据进行转录组分析,共筛选到161个差异表达基因,其中上调基因90个,下调基因71个;差异表达基因的层次聚类分析显示,ZBED6-KO猪的3个个体（spleen1、spleen3、spleen6）的表达模式相似,WT的3个个体（spleen2、spleen4、spleen5）的表达模式相似,进一步证明测序数据的准确可靠;GO和KEGG富集分析中,富集到10条显著的GO条目以及5条显著的信号通路,2条与肌肉发育相关的通路;实时荧光定量PCR试验随机检测7个差异表达基因的表达模式与RNA-seq分析结果相一致,证实了测序数据的可靠性。以巴马香猪为模型,利用RNA-seq技术研究ZBED6基因的敲除对中国地方猪脾发育的作用影响,为挖掘ZBED6基因的更多功能提供了基础。     

猪脾转移因子提高LaSota株鸡新城疫弱毒疫苗的免疫保护率

旨在了解猪脾转移因子（TF）对新城疫病毒（NDV）弱毒疫苗LaSota株的免疫增强效果及机制。本研究分别采用不同剂量（10^-2、10^-3、10^-4、10^-5羽份） LaSota疫苗株单独（单独免疫组）、LaSota疫苗株与TF联合（联合免疫组）免疫SPF鸡,14 d后以NDV F₄₈E₉强毒攻毒,同时设立对照组（非免疫攻毒）和空白组（非免疫非攻毒）,并采用ELISA方法与蛋白质芯片技术分别测定外周血IL-4、IFN-γ与IL-12 P40浓度。结果显示,鸡免疫10^-2、10^-3、10^-4、10^-5羽份时疫苗攻毒保护率和半数保护量（PD₅₀）如下：单独免疫组分别为100%、55%、0%、0%和0.000 8羽份,联合免疫组分别为100%、75%、0%、0%和0.000 5羽份,对照组和空白组死亡率分别为100%和0%。免疫10^-3羽份疫苗后,联合免疫组IL-4和IFN-γ含量均高于其他组,并于第7、14天差异极显著（P<0.01）;攻毒后,联合免疫组IL-4、IFN-γ和IL-12 P40含量均高于其他组,IL-4于第1、14天,IFN-γ于第1、3天,IL-12 P40于第1天差异极显著（P<0.01）。综上表明,TF可增强IFN-γ、IL-12介导的细胞免疫和IL-4介导的体液免疫,提高LaSota株弱毒疫苗的免疫保护率,降低疫苗PD₅₀;在抵抗NDV F₄₈E₉强毒攻击时,联合免疫组的免疫保护率明显高于单独免疫组。     

Clinical features and their course of pituitary carcinoma with distant metastasis in a dog

An 11-year-old male toy poodle with neurological symptoms was diagnosed with a macroscopic pituitary tumor, which produced adrenocorticotropic hormone. Radiation therapy with a linear accelerator was performed for the pituitary tumor, and resulted in good local tumor control. However, serum endogenous adrenocorticotropic hormone concentrations were uncontrollable even after the tumor disappeared. Abdominal computed tomography revealed splenic masses, and splenectomy was performed. Histopathological examination of the surgical specimen showed tumor cells with eosinophilic and finely granular cytoplasm suggestive of endocrine origin. Since these cells were positive for adrenocorticotropic hormone, the case was diagnosed as a pituitary carcinoma with distant metastasis. Necropsy revealed multiple metastases to the abdominal organs. This is the first case report describing canine pituitary carcinoma with distant metastasis.     

凹土纳米氧化锌对断奶仔猪脾脏抗氧化及免疫功能的影响

试验旨在探索凹土纳米氧化锌（attapulgite nano zinc oxide,ANZ）对于断奶仔猪脾脏抗氧化以及免疫功能的影响。选取体重相近的21日龄杜长大断奶仔猪210头,随机分成7组,每组6个重复,每个重复5头仔猪。分别饲喂基础日粮（CON组）、基础日粮+抗生素（ANT组）、基础日粮+3 000 mg/kg氧化锌（ZO组）、基础日粮+800 mg/kg纳米氧化锌（NZO组）和基础日粮+700、1 000、1 300 mg/kg凹土纳米氧化锌（LANZ、MANZ和HANZ组）。结果表明：与CON组相比,ZO、MANZ和HANZ组的脾脏指数显著提高（P<0.05）。与CON和ANT组相比,NZO、LANZ、MANZ和HANZ组的脾脏细胞面积显著增大（P<0.05）。与其他各组相比,MANZ组的脾脏细胞粒径显著增大（P<0.05）。与CON、ANT、NZO和HANZ组相比,MANZ组脾脏中超氧化物歧化酶（SOD）活性显著提高（P<0.05）。与NZO和ZO组相比,LANZ、MANZ和HANZ组的过氧化氢酶（CAT）活性均显著提高（P<0.05）。与ZO组相比,NZO、LANZ、MANZ和HANZ组的丙二醛（MDA）含量显著降低（P<0.05）。与CON组相比,ANT、MANZ和HANZ组的干扰素-γ（INF-γ）含量显著下降（P<0.05）。与CON、ANT、ZO和NZO组相比,MANZ和HANZ组的肿瘤坏死因子-α（TNF-α）含量显著降低（P<0.05）。与ANT组相比,MANZ组的白细胞介素-1β（IL-1β）含量显著降低（P<0.05）。与ANZ、ZO和NZO组相比,MANZ和HANZ组的IL-10含量显著增加（P<0.05）。与CON、ANT和ZO组相比,LANZ和MANZ组的IL-12的含量显著降低（P<0.05）。综上,凹土纳米氧化锌可以提高断奶仔猪脾脏的器官指数及细胞粒径、提高抗氧化功能酶的活性、减少促炎因子含量、增加抗炎因子含量,进而增强断奶仔猪的免疫以及抗氧化功能。在本试验条件下,当凹土纳米氧化锌添加量为1 000 mg/kg时效果最佳。     

四君子汤对家兔脾虚症调整作用的实验研究

为了探讨四君子汤对脾虚症的治疗作用，根据中兽医“苦寒之药损其脾胃”的理论，利用大黄煎剂复制出家兔脾虚症模型，用四君子汤治疗，结果表明：四君子汤治疗能解除脾虚症的临床症状，使实验家兔体重增加，体温回升，血清木糖吸收率，血浆胃泌素，血浆ｃＡＭＰ和ｃＡＭＰ／ｃＧＭＰ，淋巴细胞转化率，Ｅ－玫瑰花环形成率等升高，血清胆碱酯酶活性，血浆胰高血糖素降低。说明四君子汤对脾虚症有良好的调整作用。     

鸡脾脏组织抑制消减cDNA文库构建及其差异表达基因的分析

为分离与鸡抗病性密切相关的差异表达基因,并进行功能分析,利用抑制性消减杂交技术,以大骨鸡和海兰褐商品代蛋鸡20周龄时的脾脏组织为试验材料构建消减cDNA文库。文库的插入片段集中在600 bp左右,挑取760个克隆进行PCR筛选获得663个阳性克隆,经过点杂交筛选后选择了531个阳性克隆,从中随机挑取100个阳性克隆进行测序。经过同源性比对归并后得到37个差异表达基因或ESTs序列,其中,32个是已知基因,包括一般抗病性或免疫性能、特异抗病相关基因、细胞信号分子、膜蛋白质、转录因子等差异表达基因;5个为功能尚未确定的基因。并对4个可能影响鸡抗病性能的差异表达基因进行了RT-PCR半定量检测鉴定。该试验结果为进一步研究这些差异表达基因在抗病过程中的重要功能及其调控作用机理奠定了基础。     

Diagnosis and treatment of a primary splenic lymphoma in a mule

The case reported here describes a transthoracic approach for removal of a splenic lymphoma from a 5‐year‐old mule. Nine months prior to presentation at Iowa State University College of Veterinary Medicine (ISUCVM), abdominal ultrasound was performed as part of a work‐up to investigate the cause of lethargy, anorexia and anaemia. Ultrasound examination identified a haemoabdomen and multiple splenic masses. Transabdominal biopsies were collected for histopathological evaluation and lymphoma diagnosed. Based on a number of clinical and laboratory examinations there was a lack of support for multicentric involvement and all results suggested solitary splenic involvement. Surgical removal of the spleen was accomplished via a transthoracic approach without complications. Histopathology of the splenic masses revealed a B cell lymphoma. Bone marrow was collected during surgery and submitted for cytology and was found to contain a normal lymphoid population with no evidence of neoplastic cells. The mule was discharged from the hospital 11 days after surgery.     

金线莲多糖对免疫抑制小鼠脾淋巴细胞增殖及免疫器官的影响

目的：探讨金线莲多糖对免疫抑制小鼠脾淋巴细胞增殖及免疫器官的影响。方法：按大、中、小剂量给小鼠灌胃金线莲多糖,腹腔注射环磷酰胺（CY）建立免疫抑制模型;测定小鼠体重及胸腺、脾脏重量,计算胸腺、脾指数;采用MTT法检测脾淋巴细胞的增殖。结果：3个金线莲多糖剂量组小鼠的体重及免疫器官指数均低于空白组而高于模型组,脾淋巴细胞增殖均显著（P〈O．05）高于模型组。结论：金线莲多糖能提高免疫抑制小鼠体重及免疫器官指数,促进脾淋巴细胞增殖。     

Diagnosis of an Abdominal Splenosis in a Case of Ambulatory Paraparesis of the Hind Limbs in a Ferret (Mustela putorius furo)

A 2-year-old spayed female ferret (Mustela putorius furo) was presented with a 2-day history of ambulatory paraparesis of the hind limbs. On physical examination a mass was palpated in the cranial abdominal region, caudal to the stomach. A complete blood cell count revealed a normocytic normochromic anemia and reactive lymphocytes. A hypoechoic abdominal mass was observed during an ultrasound examination of the abdomen. An exploratory laparotomy was recommended and performed during which a pyramidal mass resembling a lobe of fatty liver tissue, plus omental torsion, was removed. The histopathologic diagnosis of the surgically removed mass was splenosis. The abdominal pain and ambulatory paraparesis resolved after surgery, which suggests that the paraparesis was due to pain. The findings in this report indicate that splenosis should be included as a differential diagnosis when a ferret presents with an abdominal mass and acute abdominal pain.     

USE OF TRANSSPLENIC INJECTION OF AGITATED SALINE AND HEPARINIZED BLOOD FOR THE ULTRASONOGRAPHIC DIAGNOSIS OF MACROSCOPIC PORTOSYSTEMIC SHUNTS IN DOGS

We describe the use of ultrasonography‐guided percutaneous splenic injection of agitated saline and heparinized blood for the diagnosis of portosystemic shunts (PSS) in 34 dogs. Agitated saline mixed with 1 ml of heparinized autologous blood was injected into the spleen of 34 sedated dogs under sonographic guidance. The transducer was then sequentially repositioned to visualize the portal vein, the caudal vena cava, and the right atrium through different acoustic windows. It was possible to differentiate between intrahepatic and extrahepatic shunts depending on the entry point of the microbubbles into the caudal vena cava. Portoazygos shunts and portocaval shunts could be differentiated based on the presence of microbubbles in the caudal vena cava and/or the right atrium. In one dog, collateral circulation due to portal hypertension was identified. In dogs with a single extrahepatic shunt, the microbubbles helped identify the shunting vessel. The technique was also used postoperatively to assess the efficacy of shunt closure. All abnormal vessels were confirmed by exploratory laparotomy or with ultrasonographic identification of the shunting vessel. Ultrasound‐guided transsplenic injection of agitated saline with heparinized blood should be considered as a valuable technique for the diagnosis of PSS; it is easy to perform, safe, and the results are easily reproducible.