猴头菇多糖对小鼠脾淋巴细胞体外NO分泌及iNOS mRNA表达的影响

为了研究猴头菇多糖(Hericium erinaceus polysaccharide,HEP)对小鼠脾淋巴细胞NO生成及iNOS mRNA表达的影响。采用Griess法测定HEP单独或协同ConA诱导脾淋巴细胞分泌NO含量;RT-PCR法检测脾淋巴细胞iNOS mRNA表达量。结果与细胞对照组比较,HEP在25~400μg/m L的浓度范围内能单独诱导脾淋巴细胞NO分泌及iNOS mRNA表达(P0.05);与ConA对照组比较,HEP在100~400μg/m L浓度范围内能协同Con A诱导脾淋巴细胞NO分泌及iNOS mRNA表达(P0.05)。表明HEP能单独或协同Con A诱导小鼠脾淋巴细胞分泌NO,此作用可能是通过促进i NOS mRNA表达而实现。     

沙葱黄酮对肉羊外周血淋巴细胞转化率和凋亡率的影响

本试验旨在研究饲粮中添加沙葱黄酮对肉羊外周血淋巴细胞转化率和凋亡率的影响。选用60只6月龄左右、体重(39.9±3.2)kg的小尾寒羊为试验动物,按照出生月龄和体重相近的原则,随机分为4组,每组15只羊。对照组饲喂基础饲粮,3个试验组分别在基础饲粮中添加11、22、33 mg/kg的沙葱总黄酮。预试期为15 d,正试期为60 d。正试期第60天空腹颈静脉采血,采用噻唑蓝(MTT)法测定肉羊外周血淋巴细胞转化率,采用流式细胞仪测定细胞周期、细胞凋亡率。结果表明:1)饲粮添加11、22、33 mg/kg沙葱黄酮可提高肉羊外周血淋巴细胞转化率,其中33 mg/kg组显著高于对照组(P0.05);2)与对照组相比,饲粮添加33 mg/kg沙葱黄酮能够显著提高S期和G2/M期细胞比例(P0.05);3)与对照组和11 mg/kg组相比,22和33 mg/kg组外周血淋巴细胞凋亡率显著提高(P0.05)。由此可见,饲粮中添加11~33 mg/kg沙葱黄酮可促进肉羊外周血淋巴细胞转化,并使细胞向DNA合成期转化,促进细胞分裂,对细胞凋亡有显著促进作用,以添加33 mg/kg沙葱黄酮的效果最佳。     

沙葱、油料籽实对绵羊外周血单核细胞和T淋巴细胞的影响

研究选用28只内蒙古羯羊,依照体重相近的原则随机分为4组,分别为对照组、4%沙葱组(占精料干物质的比例,以下相同)、4%沙葱+油料籽实(8%胡麻籽+2%麻籽+2%葵花籽)组、油料籽实(8%胡麻籽+2%麻籽+2%葵花籽)组,在试验期45d内采4次血,研究以上4个处理对绵羊血液细胞免疫、非特异性免疫功能的影响。结果表明:试验组单核细胞、CD2+和CD4+T淋巴细胞数均显著高于对照组(P<0.05);对照组CD8+T淋巴细胞数高于试验组,差异不显著(P>0.05)。从结果来看,沙葱和油料籽实显著提高了绵羊机体的非特异性免疫和细胞免疫机能,为以后开发舍饲肉羊新型的免疫添加剂以及进一步研究沙葱影响免疫机能的机理提供了理论依据。     

沙葱多糖提取工艺的研究

采用水提法提取沙葱多糖,试验比较了提取时间、提取温度和料液比对沙葱多糖得率的影响,并以提取时间、提取温度和料液比为考查因素,采用L(93)3正交试验确定了水提法提取沙葱多糖的最佳工艺参数,即提取时间为6h,提取温度为90℃,液料比为1:20。在上述提取条件下,水提法提取沙葱多糖得率可达到2.56%。     

黄芪多糖对小鼠腹腔巨噬细胞iNOS基因表达及NO生成的影响

研究黄芪多糖(APS)在体内对小鼠腹腔巨噬细胞诱导型一氧化氮合酶(iNOS)基因表达及产生一氧化氮(NO)的影响。分别按5、25、50、100、200mg/kg体重,连续1周以腹腔注射方式给予小鼠APS。分离小鼠腹腔巨噬细胞,QRT-PCR检测小鼠腹腔巨噬细胞中iNOS的基因表达量;培养细胞24h,硝酸还原酶法检测上清中的NO含量,培养的单层巨噬细胞做吞噬中性红试验。结果显示,APS(100mg/kg和200mg/kg)可显著地促进巨噬细胞iNOS基因的表达和NO的产生,并显著(50、100、200mg/kg)增强巨噬细胞吞噬中性红功能,APS的作用呈浓度依赖性。体内条件下APS增强巨噬细胞功能的机制之一可能是促进巨噬细胞iNOS基因的表达进而催化产生NO。     

沙葱多糖对肉羊抗氧化能力的影响

本试验旨在研究饲粮中添加沙葱多糖对肉羊抗氧化能力的影响。选择体重(30.71±2.20)kg苏尼特羯羊12只,随机分为3组,每组4个重复,每个重复1只,分别饲喂基础饲粮(对照组)、基础饲粮+0.09%(试验Ⅰ组)和基础饲粮+0.12%(试验Ⅱ组)沙葱多糖的饲粮,预试期15 d,试验期60 d。试验期第0、15、30、45、60天晨饲前颈静脉采血,分离血清;试验结束后每组随机选取3只羊屠宰,采集肝脏组织;分别测定血清和肝脏中的抗氧化指标。结果表明:与对照组相比,添加沙葱多糖均显著提高肉羊血清总超氧化歧化酶(T-SOD)、谷胱甘肽过氧化物酶(GSH-Px)、过氧化氢酶(CAT)、总抗氧化能力(T-AOC)的活性,显著降低血清丙二醛(MDA)含量(P<0.05),但试验组间差异不显著(P>0.05);由此可见,沙葱多糖能显著提高肉羊血清抗氧化指标,在一定程度上提高了肝脏抗氧化指标。     

沙葱多糖对小鼠腹腔巨噬细胞的激活作用

本试验旨在研究沙葱多糖对小鼠腹腔巨噬细胞的激活作用。采用不同浓度的沙葱多糖培养小鼠腹腔巨噬细胞,测定巨噬细胞的吞噬能力、能量代谢水平I、L-6的含量。结果表明,沙葱多糖能够增强巨噬细胞吞噬能力,提高能量代谢水平,促进IL-6的分泌。除在沙葱多糖对IL-6分泌影响的试验中,沙葱多糖浓度为400μg/mL时与空白对照组差异不显著(P0.05),其余测定指标中,试验组各个浓度均显著高于空白对照组(P0.05)。由试验结果可知,沙葱多糖对小鼠腹腔巨噬细胞具有激活作用。     

蕨麻多糖对小鼠淋巴细胞增殖和一氧化氮分泌的影响

为探讨蕨麻多糖（Potentilla anserine polysaccharide，PAP）免疫调节的作用机理，观察了PAP对小鼠脾淋巴细胞体外增殖和分泌一氧化氮的影响。结果显示，50mg／L PAP配合ConA或LPS（lipopolysaccharide，脂多糖）能使小鼠脾淋巴细胞在体外显著增殖（P〈0．05）；100、200及400mg／L PAP配合ConA或LPS可使小鼠脾淋巴细胞在体外极显著增殖（P〈0．01）；与对照组相比，PAP以不同浓度（50～400mg／L）处理小鼠脾淋巴细胞后一氧化氮分泌量均显著升高（P〈0．05）。试验结果表明，蕨麻多糖能明显促进体外培养的小鼠脾淋巴细胞增殖和分泌一氧化氮，与ConA或LPS有协同作用。     

沙葱多糖除蛋白方法的研究

试验采用L9(3)3正交试验设计,以蛋白去除率和多糖损失率为指标分别研究了Sevage法和三氯乙酸法除蛋白的最佳工艺参数,并对两种方法进行了比较。试验结果表明,Sevage法除沙葱多糖溶液中蛋白质的最佳工艺参数为:Sevage试剂添加量为沙葱多糖溶液体积的1/4,氯仿与正丁醇的比例为3:1,振摇时间为30 min。在此条件下,沙葱多糖的除蛋白率为43.82%,多糖损失率为4.67%;三氯乙酸法除沙葱多糖溶液中蛋白质的最佳工艺参数为:三氯乙酸浓度为15%,添加量是沙葱多糖溶液体积的1倍,即1:1添加,振摇时间为20 min。此时的除蛋白率为87.33%,多糖损失率为55.59%。     

阿魏根粗多糖对鸡脾脏和外周血淋巴细胞体外增殖作用的影响

为研究新疆阿魏多糖（FSP）对鸡外周血T淋巴细胞和脾脏B淋巴细胞增殖作用的影响。本研究用水提醇沉法得到阿魏总多糖（FSP_t）和3种分级多糖（FSP₃₀、FSP₅₀、FSP₇₀）,并通过苯酚-硫酸法和红外光谱对FSP进行检测。通过MTT法测定各级FSP的安全浓度及其对鸡外周血T淋巴细胞及脾脏B淋巴细胞的增殖能力。结果显示,提取的FSP均具有多糖的特征峰,为多糖类化合物。外周血T淋巴细胞增殖试验表明,除FSP₅₀外,所有多糖在一定浓度下均能单独和协同PHA刺激T淋巴细胞增殖,其中FSP_t的增殖作用较强。脾脏B淋巴细胞增殖试验表明,除FSP₅₀外,所有多糖在一定浓度下均能单独和协同LPS刺激B淋巴细胞增殖,其中FSP₇₀和FSP_t增殖作用较强。综上所述,各级FSP中,FSP_t对脾脏B淋巴细胞和外周血T淋巴细胞的增殖作用最强,可以作为进一步研究的材料。     

沙葱总黄酮对肉羊抗氧化能力的影响

本试验旨在研究基础饲粮中添加沙葱总黄酮对肉羊体内总抗氧化能力(T-AOC)、总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-PX)活力以及丙二醛(MDA)含量的影响,以阐明沙葱总黄酮对肉羊的抗氧化作用,并确定沙葱总黄酮在肉羊饲粮中的最适添加量。试验选用60只6月龄左右、体重(39.9±3.2)kg的小尾寒羊为试验动物,按照出生月龄和体重相近的原则,随机分为4组,每组15只羊。对照组饲喂基础饲粮,3个试验组分别在基础饲粮中添加11、22、33 mg/kg的沙葱总黄酮。预试期为15 d,正试期为60 d,试验期第0、15、30、45、60天时空腹颈静脉采血,分离血清,试验期结束后每组分别随机选取3只羊,进行屠宰,取肝脏和脾脏样品,分别测定血清及组织抗氧化指标。结果发现:1)饲粮添加11~33 mg/kg沙葱总黄酮可显著提高血清(第45天开始)及肝脏T-AOC(P0.05),而对脾脏T-AOC无显著影响(P0.05)。2)饲粮添加11~33 mg/kg沙葱总黄酮对血清、肝脏及脾脏T-SOD活力,血清CAT活力,血清、脾脏GSH-PX活力有一定的提高,其中33 mg/kg沙葱总黄酮的作用最强,血清中第30天后效果明显;而对肝脏、脾脏CAT活力,肝脏GSH-PX活力均无显著影响(P0.05)。3)饲粮添加11~33 mg/kg沙葱总黄酮可降低血清、肝脏MDA含量,血清MDA含量第15天后效果明显,对脾脏MDA含量无显著影响(P0.05)。结果提示,饲粮添加11~33 mg/kg沙葱总黄酮可显著提高肉羊体内抗氧化指标,并其效果具有时间和剂量依赖性,饲喂30 d后沙葱总黄酮开始发挥其体内抗氧化能力。     

Immunomodulatory effect of Artemisia argyi polysaccharide on peripheral blood leucocyte of broiler chickens

This study was conducted to investigate the immunomodulatory effect of a water‐soluble polysaccharide extracted from Artemisia argyi (AAP) in vitro. The effect was assessed in peripheral blood leucocytes (PBLs) of broilers, which were incubated with different AAP concentrations (0, 25, 50, 100, and 200 μg/ml) for 24 hr at 37°C in a 5% CO₂ incubator. The results showed that, compared with the control group, immunoglobulin M (IgM) concentration was increased in the supernatant of the 100 μg/ml AAP‐treated group (p < .05), and immunoglobulin G (IgG) concentration was increased in the supernatant of the 200 μg/ml of AAP group (p < .05). In terms of cytokine production, production of interleukin‐1beta (IL‐1β), interleukin‐6 (IL‐6) and tumour necrosis factor‐alpha (TNF‐α) in the supernatant was enhanced in the AAP group in a dose‐dependent function, as well as enhanced mRNA expressions were showed in the cells (p < .05). The highest concentration of these three cytokines was observed in different AAP groups (IL‐1β for 25 μg/ml of AAP, IL‐6 for 100, and 200 μg/ml of AAP, and TNF‐α for 100 μg/ml of AAP respectively). The concentration of nitric oxide (NO) was increased when using AAP at the concentration of 100 μg/ml (p < .05) as compared to the control group. No significant effects on inducible nitric oxide synthase, Toll‐like receptor 4 (TLR4), myeloid differentiation factor 88 and nuclear factor Kappa B (NF‐κB) mRNA level were observed at each concentration of AAP. In conclusion, we found that AAP can specifically promote the production of immunoglobulins (IgM and IgG), cytokines (IL‐1β, IL‐6 and TNF‐α), as well as the NO concentration in vitro, but not through the activation of the TLR4/NF‐κB signalling pathway.     

Effects of Allium mongolicum Regel supplementation on the digestibility,methane production,and antioxidant capacity of Simmental calves in northwest China

This study evaluated the effects of Allium mongolicum Regel (AM) supplementation on nitrogen (N) balance, ruminal fermentation, and antioxidant properties. Sixteen male calves were assigned randomly to four groups, and the four were added with 0 (CON), 200 mg/kg (body weight; BW) (Low AM; LA), 400 mg/kg (BW) (Middle AM; MA), or 800 mg/kg (BW) (High AM; HA) per day for each individual. AM was added on dry matter (DM) basis. The experiment lasted for 58 days. Supplementation of AM could significantly increase average daily gain, DM digestibility, acid detergent fiber digestibility, and retained N/Intake N. N digestibility and molar proportion of propionate in the MA and HA treatments were higher than that in the CON treatment (p < .05), respectively. AM supplementation significantly increased the molar concentration of total volatile fatty acid in the rumen fluid (p < .05). The ratio of acetate to propionate in the MA and HA groups was lower than that in the CON treatment (p < .05). Furthermore, AM supplementation significantly reduced methane (CH₄) (p < .05) emissions. AM supplementation significantly increased the activities of superoxide dismutase. The MA group could significantly increase the activities of glutathione peroxidase and decrease the content of malondialdehyde. Our results indicated that AM supplementation could affect the nutrient digestibility, CH₄ emission, and antioxidant capacity of Simmental calves.     

沙葱乙醇提取物对肉仔鸡小肠黏膜发育的影响

试验选择120只1日龄罗丝308商品肉鸡,随机分为4个处理组,每个处理组3个重复,每个重复10只鸡。对照组饲喂基础日粮,试验AWEⅠ组、AWEⅡ组和AWEⅢ组分别在基础日粮中添加100、300和500mg/kg沙葱乙醇提取物(AWE),以研究沙葱乙醇提取物对肉仔鸡小肠黏膜发育的影响。结果表明,添加500mg/kgAWE可显著提高28日龄肉仔鸡十二指肠绒毛长度及42日龄肉仔鸡空肠绒毛长度(P<0.05)。对照组及各试验组小肠绒毛宽度和隐窝深度无显著差异(P>0.05)。日粮添加沙葱乙醇提取物对肉仔鸡十二指肠V/C比值有显著提高作用(P<0.05),各试验组十二指肠V/C比值显著高于对照组(P<0.05)。28日龄时各试验组空肠V/C比值及回肠V/C比值显著高于对照组(P<0.05)。上述结果显示,在日粮中添加沙葱乙醇提取物对肉仔鸡小肠黏膜的正常发育有一定促进作用,增强了小肠的吸收能力,其中以500mg/kgAWE的添加效果最佳。     

Characterization of porcine circovirus type 2 (PCV2) infection in swine lymphocytes using mitogen-stimulated peripheral blood lymphocytes from healthy PCV2-carrier pigs

Information regarding the susceptibility of swine lymphocytes to PCV2 is rather limited. To further explore and characterize the PCV2 infection in swine lymphocytes, an in vitro model using concanavalin A (Con A)-stimulated peripheral blood lymphocytes (PBLs) obtained from clinically healthy PCV2-carrier pigs was introduced. It was found that the PCV2 antigen-containing rate was below 2% in PBLs from healthy PCV2-free pigs following treated simultaneously with Con A and PCV2. However, significantly higher PCV2 antigen- and nucleic acid-containing rates could be seen in Con A-stimulated PBLs from clinically healthy PCV2-carrier pigs. Prior to Con A treatment, both of the PCV2 antigen- and nucleic acid-containing rates in PBLs from healthy PCV2-carrier pigs were less than 1%; however, they reached 22.1+/-5.7% by flow cytometry and 27.1+/-6.5% by in situ hybridization, respectively, at 4-day post-incubation with Con A. Phenotyping of PCV2 antigen-containing cells revealed that PCV2-positive cells could be detected in both T and B lymphocyte populations within which IgM-positive B lymphocytes appeared to have a relatively higher positive rate. The Con A-stimulated PBLs also displayed a significantly higher viral load by the measurement of either PCV2 DNA copy number or viral titer when compared with the non-treated PBLs from healthy PCV2-carrier pigs. The results indicate that PBLs, especially IgM-bearing B lymphocytes, are indeed susceptible to PCV2 infection and PCV2 is capable of replicating in dividing lymphocytes. This activation-induced replication may explain in part the pathogenesis of lymphoid depletion in PMWS-affected pigs.     

藏羊血中血红素提取工艺研究

通过正交试验确定了藏羊血中提取血红素的最优工艺条件,即：溶血时添加蒸馏水1倍、氯仿5倍、分离血红素时加丙酮5倍、离心时间为20min,pH值为3。     

胸膜肺炎放线杆菌菌影疫苗免疫仔猪前后差异表达基因的鉴定与分析

为获得胸膜肺炎放线杆菌(APP)菌影诱导的仔猪淋巴细胞差异表达基因,本研究应用代表性差异分析技术构建APP菌影免疫前后正、反两个外周血淋巴细胞cDNA差减文库,并对文库中的差异基因进行克隆、测序和生物信息学分析.试验结果表明,正向文库中获得11个表达丰度上调的基因,其中7个基因与已知基因具有相似性,4个为未知新基因,经进一步功能注解发现,正向文库功能基因包括免疫信号传导相关蛋白RhoE、防御相关蛋白糖基转移样酶-1、上皮膜蛋白2、白介素-17和肿瘤免疫相关的周期素依赖性蛋白激酶抑制因子3等,这些功能基因表达丰度升高,可能有助于机体建立抗APP的免疫应答.     

日粮营养和半胱胺制剂对山羊手术前后淋巴细胞转化的影响

研究了日粮营养和口服半胱胺制剂对山羊手术前后淋巴细胞转化能力的影响。结果显示 ,正常生理状况下 ,增加日粮营养可显著提高山羊淋巴细胞转化能力 (P <0 0 5)。半胱胺制剂可改善手术创伤导致的免疫抑制 ,显著阻止严重手术创伤后淋巴细胞的刺激指数的下降 (P <0 0 5)。     

不同剂量钼对绵羊肝脏细胞的损伤作用

选用18只2月龄小尾寒羊,体质量约为（20±1.34）kg,随机分为3组,分别在基础日粮（Mo 5.61mg/kg）中添加不同剂量的钼（mg/kg）构成加钼日粮（Ⅰ组Mo 0、Ⅱ组Mo 30、Ⅲ组Mo 60）,试验周期120d,定期采血,剖杀取样,以化学比色法、流式细胞术和透射电镜的方法观察钼对绵羊肝脏细胞损伤的影响。与Ⅰ组相比：Ⅱ、Ⅲ组肝脏超氧化物歧化酶（SOD）和黄嘌呤氧化酶（XOD）活性显著下降（P〈0.01）,丙二醛（MDA）和NO含量显著升高（P〈0.01）,血清NO含量和相应抗氧化酶活性变化与肝脏中的一致;Ⅱ、Ⅲ组肝脏细胞G0/G1期和凋亡细胞百分数显著升高（P〈0.01）,S期、G2＋M期和增殖指数（PI）显著降低（P〈0.01）。与Ⅲ组相比：Ⅱ组需要更长的作用时间引起机体抗氧化酶活性、NO含量和细胞周期发生相应变化。电镜观察,Ⅱ组肝细胞出现空泡变性及线粒体肿胀,Ⅲ组细胞线粒体数目减少,严重空泡化。结论：日粮钼含量30mg/kg及其以上可引起绵羊肝脏抗氧化功能降低和NO含量升高,导致细胞增殖分化受阻,细胞受损程度与机体钼暴露剂量和时间有关。