Adipose-derived stem cells mediate immunosuppression of Th17 in multiple sclerosis

AIM: To investigate how human adipose-derived stem cells (hASCs) regulates the differentiation of Th17 cells in multiple sclerosis. METHODS: hASCs were isolated from the adipose tissues. Magnetic-activated cell sorting (MACS) kit was used to isolate CD4⁺ T cells from peripheral blood mononuclear cells (PBMCs) which were isolated by density gradient centrifugation. The percentage of CD4⁺ T cells was detected by flow cytometry. The activated CD4⁺ T cells were co-cultured with hASCs for about 4 d at different ratios of hASCs to CD4⁺ T cells (1:4 and 1:10) in a Th17 polarised condition. Another group adding anti-leukemia inhibitory factor (LIF) antibody was set up. Th17 cell proportion of the CD4⁺ T cells was determined by flow cytometry. The level of LIF in the supernatant of co-cultured system was measured by ELISA. The mRNA expression of retinoid-related orphan receptor γt (RORγt), interleukin-6 receptor (IL-6R), interleukin-23 receptor (IL-23R), LIF and leukemia inhibitory factor receptor (LIFR) was detected by real-time PCR. RESULTS: The result of flow cytometry suggested there were mainly hASCs, and the percentage of CD4⁺ T cells in the PBMCs were above 90% after MACS. The Th17 cell proportion decreased in 1:4 and 1:10 co-cultured groups in a dose-dependent manner. The mRNA expression of IL-6R, IL-23R and RORγt was downregulated and the expression of LIFR and LIF was up-regulated. When the anti-LIF was added into the co-cultured system, the ratio of Th17 cells increased and reached to the control level. The protein level of LIF obviously increased after co-cultured. After anti-LIF added, the mRNA expression of RORγt and IL-6R was up-regulated. CONCLUSION: hASCs inhibits the differentiation of Th17 cells from multiple sclerosis patients through the competitive inhibition of LIF/IL-6 by secreting LIF.     

Regulatory effects of spirolactone on SiO2-induced lung macrophage subtype switching

AIM: To investigate the influence of spirolactone (SPI) on mouse pulmonary macrophage subtype switching induced by silicon dioxide (SiO₂). METHODS: C57BL/6 mice were divided into normal saline (NS) group, SiO₂ group, SiO₂+SPI group and SiO₂+NS group. A mouse model of silicosis was developed with crystalline SiO₂ particles (40 mg/kg, via oropharyngeal instillation). SPI (20 mg/kg) or (NS) was delivered daily by oral gavage after SiO₂ administration. The mice were sacrificed on days 3, 14 and 28. Alveolar washing, total cell counting, differential cell counting and flow cytometry analysis were performed. The right lower lobe of lavaged lungs was collected to prepare single-cell suspension for flow cytometry analysis. RESULTS: Compared with SiO₂ group, SPI treatment significantly alleviated SiO₂-induced inflammatory cell accumulation in brochoalveolar lavage fluid on day 3 and 14. Compared with NS group, the M1 alveolar macrophages and M1 pulmonary interstitial macrophages in SiO₂ group switched to M2 subtype dramatically, while SPI treatment reversed the switching effectively.CONCLUSION: SPI treatment alleviates SiO₂-induced inflammatory cell accumulation, which may be related to reversing macrophage subtype switching.     

梅花鹿IFN-α基因的克隆与原核表达研究

为研究梅花鹿IFN-α的抗病毒功能和分子机制,根据GenBank中牛IFN-α基因序列(A00145),设计并合成1对引物,以梅花鹿肝脏组织DNA为模板,采用PCR技术扩增梅花鹿IFN-α全基因核苷酸序列。序列分析表明梅花鹿IFN-α基因全长570 bp,编码189个氨基酸,其中含18个氨基酸的信号肽和169个氨基酸的成熟多肽,存在1个潜在跨膜区,基因随不同物种发育进化地位表现出种属间的差异性。在此基础上,合成1对引物,扩增梅花鹿IFN-α成熟肽核苷酸序列,构建pET28a-IFNα原核重组表达质粒并转化至大肠杆菌BL21(DE3)感受态细胞中。SDS-PAGE和Western blotting检测结果表明,在24 kd左右出现特异性蛋白带,表达产物主要为不溶性的包涵体,表达量占菌体总蛋白的34.04%,且表达产物与抗His标签抗体可发生特异性反应。将Ni柱纯化的pET28a-IFNα诱导表达产物复性后,细胞病变抑制法检测表明表达产物具抗病毒活性。研究结果为梅花鹿IFN-α蛋白抗病毒分子机制的研究及抗病毒药物的开发奠定了基础。     

广东鲂全人工繁殖的研究

本文研究了广东鲂人工繁殖的鱼苗在池塘育成亲鱼的方法,用ＬＲＨ－Ａ２与ＤＯＭ混合催们效果最优,人工授精的关键是效应时间的掌握,本研究提出广东鲂最小性成熟年龄为２龄,属一年我次产卵鱼类,可进行一次２次性成熟产卵,胚胎发育程序与家鱼基本相似,胚胎发育最适水温范围约为２４－２８℃,临界水温上、下限分别为３１℃和２０℃。     

万寿菊茎段组织培养

本文以万寿菊的茎段为外植体,将其接种在附加不同浓度的 Ｂ Ａ 和 Ｎ Ａ Ａ 的 Ｍ Ｓ 培养基中进行培养。结果表明： Ｍ Ｓ＋ Ｂ Ａ Ｏ．１ ＋ Ｎ Ａ Ａ Ｏ．１ 是万寿菊茎段培养适宜的培养基,诱芽率和生根率分别为９３ ．３ ％ 和９０ ％ ,并可１ 次成苗。     

陕西省重大动物疫病秋防免疫抗体的检测与分析

猪瘟、口蹄疫、禽流感和新城疫四种重大动物疫病对畜禽业的危害非常严重,目前疫苗免疫是预防和控制动物疫病最有效的途径,免疫抗体检测能很好的反映出免疫效果的好坏。为了及时了解陕西省重大动物疫病免疫状况,2010年11月份在全省11个市(区)的21个县随机采集牛、羊、猪、鸡秋防免疫血清共2359份,按照农业部规定的检测方法,对其进行了重大动物疫病免疫抗体水平的检测,检测结果均达到或超过农业部规定的合格标准,说明陕西省今年秋防动物免疫效果总体较好。     

水生生物近缘种和产地的分子生物学判别

随着分子生物学技术的发展,渔业生物近缘种及产地的判别由形态学深入到了蛋白质和DNA水平。本文综述了目前已经应用于水生生物的相关分子生物学技术（包括色谱、免疫学判别以及蛋白的电泳、DNA的序列分析、PCR-RFLP、RAPD、AFLP、SNP、SSRI、SSR、Real-Time PCRs、pecies-specific primer PCR和PCRlab-on-a-chip等）以及它们各自的优缺点;在把握了这些研究进展的基础上,展望了相关技术的发展趋势。     

刺参呼吸树抗氧化防御酶类基因在夏眠期的表达特征

以夏眠刺参呼吸树为研究材料,研究了刺参抗氧化防御酶类的基因表达特征,共获得夏眠刺参的5个基因片段序列:铜锌超氧化物歧化酶(Cu/Zn-SOD)、过氧化氢酶(CAT)、磷脂氢谷胱甘肽过氧化物酶(PHGPx)、过氧化物还原酶5(PRDX5)及过氧化物还原酶6(PRDX6),完成基因序列初步分析;利用荧光定量PCR技术,以NDUFA13 (NADH dehydrogenase［ubiquinone］1 alpha subcomplex subunit 13)和 ACTB (beta actin)基因为多内参,定量分析实验各阶段刺参抗氧化防御酶类基因表达量。结果表明,刺参抗氧化酶类基因与近、远源物种同源性均较高;现场调查实验结果显示,相对5月非夏眠刺参样品,8月夏眠刺参呼吸树组织Cu/ZnSOD及PRDX5基因表达上调,CAT及PHGPx基因表达量均出现显著下降。温度诱导夏眠实验证实Cu/ZnSOD基因在实验第0天表达量显著上升至2.01倍,并一直保持较高水平;CAT基因表达在实验第0天下调,第5天时出现上调,而在第10天和40天表达量均出现显著下降;PHGPX基因表达实验期间均出现显著下调,在第40天时降到最低,为0.34倍;PRDX5基因表达在实验第0天出现显著上调至1.55倍,第5天回落后呈现上升,于第40天达到高值1.91倍;PRDX6基因表达在实验第10、20天出现显著下调。     

壳聚糖絮凝法回收鱼糜漂洗水中水溶性蛋白质的工艺研究

采用壳聚糖作为絮凝剂进行鱼糜漂洗水中水溶性蛋白质的回收试验。分别通过体系的pH、回收温度和壳聚糖絮凝剂加入量的单因素试验及这3个因素的正交试验,研究其对蛋白质回收率、透光率和化学耗氧量（COD）去除率的影响,得出壳聚糖絮凝法回收40mL已知蛋白质质量浓度（9．0mg·mL-^1）的鱼糜漂洗水中水溶性蛋白质的最佳工艺条件为体系pH8．0,回收温度35℃,1％壳聚糖添加量为1．5mL。该工艺获得的蛋白质回收率为73．17％,透光率87．50％,COD去除率47．20％。     

基于BP神经网络模型的福建海域赤潮预报方法研究

赤潮往往给渔业生产和人类的生命安全造成极大的危害,但由于赤潮的成因十分复杂,对其进行预报非常困难。本研究收集了福建海区2000年至2016年发生的219个赤潮案例有效数据,应用BP神经网络人工智能模型建立了其与气温、降水、风速、气压和日照5个气象因子的非线性关系,并将这些赤潮案例数据与相应的气象指标按闽东、闽中和闽南3个海区,分别输入模型进行学习、训练与预测。结果显示:1)闽东海区53个训练样本45个预测正确,正确率达84.91%,3个模拟预测样本全部正确;2)闽中海区69个训练样本58个预测正确,正确率达84.06%,4个模拟预测样本全部正确;3)闽南海区85个训练样本的运算预测结果63个正确,正确率74.12%,5个模拟预测样本全部正确,达到预期的结果。研究表明,以气象因子为自变量采用BP神经网络模型对赤潮的发生进行预测是可行的,该方法可为赤潮的预测提供新的途径。