牛分枝杆菌MarP蛋白的表达纯化及其单克隆抗体制备

为制备能够特异性识别牛分枝杆菌（M.bovis）抗酸蛋白酶（MarP）的特异性单克隆抗体（MAb）,本研究利用大肠杆菌表达系统表达了MarP蛋白的胞浆区,并以β-casein为底物、DTT为抑制剂检测MarP的丝氨酸蛋白酶活性。以具有活性的MarP蛋白免疫小鼠,取脾细胞与SP2/0细胞融合,并加入饲养层细胞轻轻摇匀分装于96孔培养板,置于37℃、5% CO₂培养箱内培养10 d,通过IFA和间接ELISA筛选阳性细胞株进行亚克隆,制备能分泌针对MarP MAb的杂交瘤细胞株,并通过Western blotting和ELISA检测MAb与重组表达和天然的MarP蛋白的反应性。结果显示,重组表达的MarP具有良好的丝氨酸蛋白酶活性,能够在12 h内将30 μg的β-casein酶解完全,DTT能够抑制MarP的酶活性。具有活性的MarP蛋白免疫小鼠后,获得5株针对MarP的MAb,均能够特异性识别重组和牛分枝杆菌天然表达的MarP蛋白的线性表位,而不与大肠杆菌（E.coli）和耻垢分枝杆菌（M.smegmatis）的蛋白反应。本研究成功制备了MarP蛋白及MAb,为进一步研究MarP的作用底物及其在牛分枝杆菌抗酸胁迫中作用机制提供了必备材料。     

巴戟枯萎病的病原菌鉴定

 巴戟枯萎病是一种毁灭性的病害,且前在广东省高要县发生严要.经病原菌分离培养、形态特征观察和致病性测定等试验结果表明,它由尖镰孢霉内的一个新的专化型(Fusarium oxysporum Schl f.sp.morindae Chi et Shi f.sp.nov.)为害所致.病菌可通过土壤和插条传播.据初步研究表明,用50%多菌灵可湿性粉刺1000倍浸泡液插条12—24小时可防治带菌,值得继续试验.     

Effects of gastrin on expression of cyclooxygenase and growth factors in rat gastric mucosa

AIM: To clarify the effects of gastrin on the expression of cyclooxygenase (COX) and several growth factors in rat gastric mucosa. METHODS: Male Sprague Dawley rats were fasted for 24 hours and subcutaneously injected with saline or gastrin 17 at doses of 1 μg/kg, 10 μg/kg and 100 μg/kg, respectively. The expression of COX-1, COX-2, heparin-binding epidermal growth factor-like growth factor (HB-EGF) and hepatocyte growth factor (HGF) in the gastric mucosa were examined using Western blotting and immunohistochemical staining. Effects of a potent gastrin receptor antagonist YM022 on the expression of COX-1, COX-2, HB-EGF and HGF in gastric mucosa were also evaluated. RESULTS: Gastrin dose-dependently increased the expression of COX-2 and HB-EGF in rat gastric mucosa while the expression of COX-1 and HGF did not change significantly after treatment with gastrin. However, pretreatment with YM022 dose-dependently abolished the up-regulation of COX-2 and HB-EGF expression induced by gastrin. CONCLUSIONS: This study demonstrates that gastrin up-regulates COX-2 and HB-EGF expression in rat gastric mucosa, indicating that COX-2 and HB-EGF are involved in pathogenesis of the gastrin-related gastric mucosal hyperplasia and carcinoma of stomach.     

三明市区家鼠寄生革螨的调查报告

本文报告了三明市区家鼠寄生革螨的种类、组成、季节消长情况。观察一周年，共捕鼠７１１只，带螨鼠１４３只，带螨率为２０．１１％，检集革螨８７１只，鼠革蛲指数为１．２３只／鼠，鉴定为３属４种即毒厉螨、纳氏厉螨、柏氏禽刺螨和鼠颚毛厉螨；毒厉螨为优势种，占８１．８６％，平均螨指数为１．００只／鼠，各宿主动物间带情况存在差异。以褐家鼠最高，达３７．２０％，螨指数为２．７２只／鼠，革螨常年活动，以４－６月、８３     

福安特抗小鼠移植性肿瘤的实验研究Ⅰ

目前，肿瘤预防研究的主要研究目标之一是寻找高效、低毒的药物。作为福安特注射液主要成分的熊果酸，1990年就被日本列为最有希望的肿瘤预防药物之一。熊果酸（ursolic acid，UA）又名乌索酸，乌苏酸，属三萜类化合物，抗肿瘤活性是熊果酸最主要的药理作用，同时也具有其他广泛的生物活性，     

灵芝和菌糠降低鸡蛋胆固醇的试验研究

选取48周龄海兰褐商品蛋鸡120羽,随机分成3组,试验期为4周,对照组Ⅰ组喂基础日粮,试验Ⅱ组添加2.5g/kg灵芝,试验Ⅲ组添加2.5g/kg菌糠,在一致的背景下,探讨灵芝、菌糠对鸡蛋胆固醇的影响。结果表明:Ⅱ、Ⅲ组可降低蛋鸡血液甘油三酯(TG)、总胆固醇(TC)含量,增高血清高密度脂蛋白胆固醇(HDL-C)含量;3组之间料蛋比差异显著,Ⅱ、Ⅲ组料蛋比分别比对照组低6.4%(P<0.05)、11.0%(P<0.05)。Ⅱ、Ⅲ组平均产蛋率分别比对照组提高5.38%(P<0.05)和3.29%(P<0.05)。试验4周,Ⅱ、Ⅲ组鸡蛋胆固醇含量分别降低15.3%(P<0.05)和10.4%(P<0.05),且两组间差异显著(P<0.05),研究表明灵芝及菌糠均能提高蛋鸡生产性能,降低鸡蛋胆固醇含量。相比之下,灵芝是降低鸡蛋胆固醇较为理想的添加剂。     

27-Hydroxycholesterol modulates lung cancer cell proliferation by activation of LXR signaling pathway

AIM:To investigate the effect of cholesterol metabolite 27-hydroxycholesterol (27-OHC) on the proliferation of lung cancer cells. METHODS:Human lung cancer A549 cells were treated with 27-OHC at different concentrations (0, 0.3125, 0.625, 1.25, 2.5, 5 and 10 μmol/L) for 24~48 h. The cell viability, cell cycle, cell prolife-ration, the intracellular cholesterol levels and cholesterol metabolism-related molecule expression were subsequently assessed by CCK-8 assay, flow cytometry, EdU staining, tissue total cholesterol detection kit, real-time PCR and Western blot. RESULTS:27-OHC decreased the viability of the A549 cells in a dose-and time-dependent manner (P<0.01) and inhibited the cell proliferation (P<0.05). The expression of typical liver X receptor (LXR) downstream target proteins including ATP-binding cassette transporter A1 (ABCA1), low-density lipoprotein receptor (LDLR), and 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMG-CR) were modulated, which promoted the efflux of intracellular cholesterol, and reduced cholesterol influx and de novo synthesis, resulting in decreased intracellular cholesterol levels and cell viability. Furthermore, the inhibitory effect of 27-OHC on A549 cell viability was significantly attenuated after the LXR pathway was partially blocked by 5 μmol/L GSK2033 treatment (P<0.05). CONCLUSION:27-OHC inhibits A549 cell prolife-ration via activation of LXR signaling pathway.     

小檗碱对炎症性肠病肠道屏障的影响及其信号通路

小檗碱是传统中药黄连等植物的主要活性成分,具有抗菌消炎、调节机体免疫等作用,可改善由炎症性肠病(IBD)导致的肠道屏障损伤。小檗碱通过抑制Toll样受体(TLRs)/核转录因子κB(NF⁃κB)、Janus活化激酶(JAK)/信号转导与转录激活子(STAT)信号通路和激活胰岛素样生长因子1(IGF⁃1)/胰岛素样生长因子结合蛋白(IGFBPs)信号通路,缓解IBD引起的肠道机械屏障、化学屏障、免疫屏障和微生物屏障功能损伤。本文就小檗碱的来源及理化性质、IBD的病理特征、小檗碱对IBD肠道屏障的修复作用及其信号通路进行综述,以期为畜牧生产中保护动物肠道健康提供参考。     

马铃薯晚疫病菌对甲霜灵抗性的快速检测方法

为快速检测马铃薯晚疫病菌——致病疫霉Phytophthora infestans对甲霜灵的抗性,基于已知致病疫霉RPA190基因的T1145A变异引起的F382Y氨基酸点突变与甲霜灵抗性有关,通过设计4对特异性引物F382Y-F1/F382Y-R、F382Y-F2/F382Y-R、F382Y-F3/F382Y-R和F382Y-F4/F382Y-R建立等位基因特异性PCR(allele specific-polymerase chain reaction,AS-PCR)快速分子检测法,其中F382Y-R引物在4对引物里保持不变,并分析所建分子检测法的特异性和灵敏度。结果表明,正向特异性引物F382Y-F2、F382Y-F3和F382Y-F4的3''末端在致病疫霉抗甲霜灵菌株原有核苷酸突变位点1145A(对应引物为F382Y-F1)的基础上,再人工引入第1144位的核苷酸突变,将1144T分别突变成1144G、1144C和1144A,并优化退火温度和特异性引物与内参引物ITS1/ITS4比例,最终确定最适退火温度分别为54、60和58℃,特异性引物与内参引物最适浓度比例均为5∶1。以该3条引物对应的3对特异性引物和内参引物ITS1/ITS4组成的3组多重AS-PCR引物对甲霜灵敏感和抗性菌株具有良好的特异性,敏感菌株的扩增产物含1条879bp的内参片段,抗性菌株的扩增产物为1条879bp的内参片段和1条461bp的目的片段。3组AS-PCR检测体系均具有较高的灵敏度,其中引物F382Y-F4/F382Y-R对致病疫霉DNA的检测灵敏度达0.4pg/μL,引物F382Y-F2/F382Y-R和F382YF3/F382Y-R的检测灵敏度达4pg/μL。     

桐柏大枣组织培养育苗技术研究

用组织培养法培育桐柏大枣苗，适宜的诱导培养基为MS 6-BA2.0mg/L KT0.5mg/L 2,4-D0.2mg/L 3%蔗糖，生根培养基为MS 6-BA0.01mg/L IBA1.0mg/L IAA0.5mg/L 3%蔗糖，用幼树枣头顶芽制备外植体效果最好。