Metastatic immune infiltrates correlate with those of the primary tumour in canine osteosarcoma

Our lack of understanding of the immune microenvironment in canine osteosarcoma (cOSA) has limited the identification of potential immunotherapeutic targets. In particular, our ability to utilize readily available tissue from a dog's primary tumour to predict the type and extent of immune response in their pulmonary metastatic lesions is unknown. We, therefore, collected 21 matched pairs of primary tumours and pulmonary metastatic lesions from dogs with OSA and performed immunohistochemistry to quantify T‐lymphocyte (CD3), FOXP3+ cell, B‐lymphocyte (Pax‐5), and CD204+ macrophage infiltration. We found that T‐lymphocytes and FOXP3+ infiltrates in primary tumours positively correlated with that of metastatic lesions (ρ = 0.512, P = 0.038 and ρ = 0.698, P = 0.007, respectively), while a strong trend existed for CD204+ infiltrates (ρ = 0.404, P = 0.087). We also observed T‐ and B‐lymphocytes, and CD204+ macrophages to be significantly higher in a dog's pulmonary metastasis compared to their primary tumour (P = 0.018, P = 0.018, P = 0.016, respectively), while FOXP3+ cells were only significantly higher in metastases when all primary tumour and metastasis lesions were compared without pairing (P = 0.036). Together, these findings suggest that the metastatic immune microenvironment may be influenced by that of the primary cOSA, and that primary tumour immune biomarkers could potentially be applied to predict immunotherapeutic responses in gross metastatic disease. We, therefore, provide a rationale for the treatment of cOSA pulmonary metastases with immunotherapeutics that enhance the anti‐tumour activity of these immune cells, particularly in dogs with moderate to high immune cell infiltration in their primary tumours.     

西北干旱灌区紫花苜蓿高产田施肥效应及推荐施肥量研究

为揭示紫花苜蓿氮、磷、钾肥效应,采用“3414”不完全正交回归设计,对紫花苜蓿氮、磷、钾肥合理配比施肥效应进行研究,同时对紫花苜蓿产量及蛋白总量进行肥效模型拟合。结果表明,氮、磷、钾对建植2年苜蓿产量的贡献为钾>磷>氮,对建植3年苜蓿产量的贡献为磷>钾>氮,建植2与3年苜蓿交互效应均表现为氮磷>氮钾>磷钾。氮、磷、钾对建植2年苜蓿蛋白总量的贡献为氮>钾>磷,对建植3年苜蓿蛋白总量的贡献为氮>磷>钾。建植2年苜蓿氮磷肥互作效应明显优于氮钾、磷钾互作;建植3年苜蓿氮磷、氮钾交互对苜蓿蛋白总量的增产效果明显大于磷钾交互。采用频度分析法,通过模拟寻优,得出建植2年紫花苜蓿目标产量大于平均产量17522kg·hm^-2时,优化施肥量为氮56.27~67.51kg·hm^-2、磷77.69~90.48kg·hm^-2、钾76.43~87.18kg·hm^-2;建植3年紫花苜蓿目标产量大于平均产量19234.1kg·hm^-2时,优化施肥量为氮46.75~57.66kg·hm^-2、磷80.15~92.28kg·hm^-2、钾57.79~69.74kg·hm^-2;建植2年紫花苜蓿目标蛋白总量大于平均2115kg·hm^-2时,优化施肥量为氮66.35~77.48kg·hm^-2、磷79.34~92.87kg·hm^-2、钾73.68~85.38kg·hm^-2;建植3年紫花苜蓿目标蛋白总量大于平均2656kg·hm^-2时,优化施肥量为氮68.44~79.50kg·hm^-2、磷72.74~85.96kg·hm^-2、钾50.68~61.61kg·hm^-2。     

用RAPD标记分析高羊茅的遗传多样性

采用RAPD分子标记技术对从国外引进的15个高羊茅品种的遗传多样性进行了研究。从60个随机引物中筛选出12个有效引物,它们共扩增出85条DNA带,其中59条为多态性带,占69.41%,平均每个引物扩增出多态性带4.92条;利用NTSYS-PC软件计算出的不同品种间Jaccard遗传相似系数(GS)变化范围较大,为0.373~0.932;根据得到的遗传相似性矩阵进行非加权组法(UPGMA)聚类分析,建立高羊茅品种的分子系统树状图;以相似系数0.68为标准,可将所有品种分为3类,品种翠丽和贝克各自聚为一类,其余13个品种聚成一类。     

鄱阳湖区血吸虫病流行病学调查与疫情分析

为了调查湖区血吸虫病流行环境与传染源构比,找出主要传染源、传播途径和场所,进一步揭示流行规律,为制定血防对策提供依据.调查范围为鄱阳县及两乡四村.结果,草洲钉螺占99.4%,90%以上牛上洲放牧,人粪流放率31%,其中渔民78%,牛粪流放率78%,其中64%直接污染草洲水体,野粪构比中牛占92.8%,传播粪量牛占98.85%,人占0.96%,其中渔民占0.12%.结论认为湖区疫情重,主要传染源是牛,主要感染途径是接触疫水,疫水形成的主因是牛上洲放牧,主要感染场所是草洲及周边水体.增加血防投入,稳定队伍,实施查治病和＂四个突破＂为主体的防治策略,控制畜源性传染源,净化环境与草洲,切断传播途径,才能有效控制血吸虫病.     

猪细小病毒NS基因原核表达与多克隆抗体制备

旨在制备抗猪细小病毒(PPV)非结构蛋白共有氨基酸的NS多肽多克隆抗体。根据GenBank(MK993540)公布的PPV基因组序列,克隆其非结构蛋白NS1、NS2共有基因序列(NS基因),并进行生物信息学分析。进而将NS基因克隆至原核表达载体pET-32a,构建重组质粒pET-32a-NS,转化至大肠埃希菌Rosetta(DE3)中进行诱导表达,利用镍柱亲和层析技术纯化表达的重组多肽,用重组多肽免疫Balb/c小鼠制备多克隆抗体。结果显示,PPV杨凌株NS基因长258bp,编码86个氨基酸的多肽,是具有亲水性的非跨膜NS多肽,具有大量的B细胞线性表位。NS多肽在37℃、0.8mol/L IPTG条件下,诱导6h有大量的可溶性表达。免疫印迹试验结果显示,该多肽具有较好的抗原性。用纯化NS多肽免疫小鼠后获得鼠抗NS多肽的血清抗体效价为1∶12800。用制备的NS多克隆抗体检测纯化的NS重组多肽及真核表达的NS1蛋白,均能检测出相应特异性条带,为进一步研究NS蛋白在PPV致病过程中的作用奠定了基础。     

Effect of vehicular volume on the early pulmonary injury and inflammatory response in rats inoculated intratracheally with silica

The effect of vehicular saline solution volume on early lesions induced in rats by intratracheal administration of silica was evaluated. Seventy-two male Long-Evans rats were randomly assigned 6 each to 12 factorial groups (3 X 2 X 2): 3 doses of silica (0, 2.5, and 5 mg), 2 volumes of vehicle (saline solution; 0.1 and 0.5 ml), and 2 postinoculation times (1 and 3 days). Lactate dehydrogenase and alkaline phosphatase activities in the bronchoalveolar lavage fluid supernatant and cell viability of bronchoalveolar cells were used as indicators of cell injury. The number of pulmonary alveolar macrophages and polymorphonuclear leukocytes were used as indicators of inflammatory response. Dose of silica and postinoculation time had a significant (P less than 0.05) effect on the biochemical and cellular composition of lavage fluid. The volume of vehicle in which silica was suspended significantly (P less than 0.05) enhanced the pulmonary injury and inflammatory response. However, dose-volume interaction was only significant (P less than 0.05) in 1 of 6 parameters, indicating that the effect was additive, but not synergistic, in nature. Seemingly, vehicle volume had an enhanced effect on the injury and the inflammatory response induced by intratracheal inoculation of silica.     

一起输入性羊痘疫情的暴发调查

2022年2月云南省元江县某农户饲养的山羊出现疑似羊痘病情,袭击率为16.91%(46/272),病死率为45.65%(21/46)。为确定病因,防止病情扩散,开展了紧急流行病学调查。综合病羊临床症状以及现场调查和实验室检测结果,确诊本起疫情为输入性羊痘疫情。通过采取病羊与健康羊群隔开饲养、紧急免疫、对症治疗、消毒、加强饲养管理等干预措施,病情得到有效控制。分析认为,养殖户疫病防控意识淡薄,缺乏生物安全防范意识,调入携带病原羊只混群饲养等是引发病情的主要原因。本次病情警示,养殖场应加强生物安全管理,做好未出售羊群返场的隔离检疫,同时要加强饲养管理,减少应激因素,以预防疫病发生。     

低浓度阿奇霉素对猪链球菌2型蛋白表达、荚膜多糖与药物敏感性的影响

本研究通过探索低浓度阿奇霉素对猪链球菌2型蛋白表达、荚膜多糖与药物敏感性的影响,为进一步研究环境中低浓度抗生素对微生物的影响奠定基础。用低浓度阿奇霉素处理猪链球菌2型,利用蛋白质组学iTRAQ技术,筛选关键差异表达蛋白。同时测定猪链球菌2型的荚膜多糖含量和药物敏感性。结果显示,共鉴定到差异表达蛋白174个,占总鉴定蛋白的11.6%,多数差异表达蛋白参与催化和代谢过程,属于膜蛋白,其中,3个荚膜多糖蛋白、9个ABC转运蛋白、1个50 S核糖体蛋白、1个核糖体RNA甲基转移酶、1个DNA回旋酶上调表达;8个荚膜多糖蛋白、4个50S核糖体蛋白、4个30S核糖体蛋白、1个核糖体RNA甲基转移酶、1个DNA聚合酶IV下调表达。用低浓度阿奇霉素处理猪链球菌2型,发现其对多种抗生素的敏感性降低,存活下来的菌株,恢复正常培养后,药物敏感性恢复。用低浓度阿奇霉素处理猪链球菌2型,荚膜多糖含量也未发生大幅度变化。猪链球菌2型通过改变自身蛋白质组的表达量,以适应低浓度阿奇霉素的选择压力。与低浓度阿奇霉素共存时,猪链球菌2型开启外排泵系统,会增加细菌产生多重耐药的风险。