铺散诸葛菜的组织培养及植株再生

 以铺散诸葛菜的叶柄及叶片为外植体, 研究了不同的再生方式。在6-BA 和NAA 配合使用的MS 培养基上, 叶柄和叶片外植体有高的成芽率, 具有强的器官再生能力。在2,4-D 与6-BA、NAA 结合的培养基上, 叶柄和叶片有较高的愈伤组织诱导率, 在含6-BA 的培养基上也易于分化。叶柄、叶片再生的芽和经愈伤组织分化而得来的芽快速繁殖后, 在生根培养基上形成完整植株。研究表明铺散诸葛菜具有极强的器官分化能力。     

黄瓜前中期染色体C - 分带及其制备方法研究

 以活体种子根为材料, 采用放线菌酮预处理, 直接获得了清晰的黄瓜前中期染色体C - 分带。黄瓜前中期染色体的长度介于3～8μm , 单套染色体组具有21 条稳定的C - 带, 包括12 条末端带、7 条着丝点带、1 条中间带和1 条随体带。还探讨了影响前中期C - 分带制备的关键因素, 提出了黄瓜前中期染色体C - 分带的实用制备方法。     

茶薪菇原生质体制备及诱变的研究

对茶薪菇原生质体的分离及再生进行研究 ,结果表明以溶壁酶的去壁效果最好 ,不同酶组合可提高去壁效果 ,酶解液在 2 0 %时原生质体释放量很高。酶解适宜温度为 30℃ ,培养 4～ 5d的菌丝酶解 3h释放的原生质体最多。以甘露醇作为渗透压稳定剂 ,原生质体的释放量及再生率均较高 ,再生率达到 5 %左右 ,经紫外线照射 2 0s,致死率即达 70 %以上     

Model establishment and biological behaviour observation of mouse blastocyst co-cultured with hepatocarcinoma cell lines with differently invasive and metastatic potential in vitro

AIM: To explore interaction and biological behaviour changes of two kinds of cells-blastocysts and hepatocarcinoma cells in the same microenvironment. METHODS:The models of mouse blastocysts co-cultured with human hepatocarcinoma cell lines were established, then biological behaviours and mutual effects of the two kinds of cells in co-culture system were observed. RESULTS: Compared with control group, hepatocarcinoma cells with differently invasive and metastatic potential significantly enhanced the rates of blastocyst hatchment , attachment and outgrowth(P＜0.05). There was no significant difference in those among hepatocarcinoma cells co-cultured groups (P＞0.05). The blastocyst hatched and attached to hepatocarcinoma cells with differently invasive and metastatic potential. Then, differential trophoblasts invaded hepatocarcinoma cells. The clear-cut interfaces were gradually formed between both sides. Hepatocarcinoma cells on interface showed changes of growth direction and cell shapes and did not invade blastocysts. CONCLUSIONS: Hepatocarcinoma cells promoted blastocyst development. Blastocysts implanted and invaded hepatocarcinoma cells with differently invasive and metastatic potential in vitro, which indicate that blastocyst implantation in vitro does not relate with the kinds and differential level of interactional cells and the low selectivity maybe relate with high adaptability of early life.     

Nitric oxide-mediated the cardioprotection of tumor necrosis factor-alpha on cultured neonatal rat cardiomyocytes during hypoxia/reoxygenation

AIM: To investigate the role of nitric oxide synthase (NOS), soluble guanylyl cyclase (sGC) and protein kinase C (PKC) signaling in tumor necrosis factor-α (TNF-α)-induced cardioprotection against hypoxia/reoxygenation (H/R) injury. METHODS: Neonatal rat ventricular myocytes were pretreated with TNF-α or sodium nitroprusside (SNP) or L-arginine (L-Arg), respectively, for 12 h and then subjected to continuous hypoxia for 12 h, followed by reoxygenation for 6 h. The manganese superoxide dismutase (Mn-SOD) activity of the cells was measured after H/R. Myocyte injury was determined by the release of lactic dehydrogenase (LDH). RESULTS: TNF-α (10⁵ U/L) significantly increased the Mn-SOD activity and decreased release of LDH from ventricular myocytes. The cardioprotection against H/R injury was induced by the pretreatment with SNP (5 μmol/L) or L-Arg (5 mmol/L), which was blocked by ODQ (10 μmol/L), the specific sGC inhibitor, and Chel (5 μmol/L), the specific PKC inhibitor. Pretreatment with L-NAME (100 μmol/L), ODQ, Chel, antoxidant 2-MPG (400 μmol/L) or tyrosine kinase inhibitor genistein (50 μmol/L) attenuated the increased Mn-SOD activity and reduced LDH level induced by TNF-α. CONCLUSION: The results suggest that NO may play a role in TNF-α-induced cardioprotection, which is mediated by sGC and PKC.     

Effects of β-ME and RA on expression of GFAP in mesenchymal cells derived from mouse fetal liver

AIM: To investigate the effects of β-mercaptoethanol (β-ME) and all-trans rentinal acid (RA) on glial fibrillary acidic protein (GFAP) expression in mesenchymal cells derived from mouse fetal liver in vitro. METHODS: Cells suspension from 14.5-days-old mouse fetal liver were cultured in DMEM/HEPES/F12 supplemented with 20％ FCS and mesenchymal cells were acquired after discarding nonadherent cells. The 5th passage cells were induced by β-ME and RA. The characteristics of treated cells were assayed by immunocytochemistry staining at 5 hours and 5 days after induction. β-actin as an internal control, GFAP gene expression of mesenchyal cells was detected with semi-quantitative RT-PCR. RESULTS: After being inducted by β-ME and RA, 80％ approximately of the cells exhibited typical neural morphology and about 85％ expressed GFAP phenotype. Semi-quantitative RT-PCR showed that mRNA expression of GFAP increased in treated cells versus untreated cells (P<0.01). CONCLUSION: GFAP expression in mesenchymal cells derived from mouse fetal liver in vitro increases after being treated with β-ME and RA.     

秦巴山区养羊户适度经营规模的研究

通过对秦巴山区陕西省西乡县15个典型陕南白山羊养殖专业户的资金投入、羊群结构、草地资源、经济效益等情况的调查，分析了养羊户的资源利用和生产经营状况，确定了与山羊生产密切相关的12个变量，即种公羊数、基础母羊数、后备母羊数、育肥羊数、羊舍面积、草地补种及施肥费用、医杂费、氨化或青贮饲料量、补饲精料量、工作日、投资额，并确定了各变量的参数值，以利润最大为目标函数，然后运用线性规划模型，计算出不同投资、不同人力及草地资源等条件下的适度经营规模。结果表明，养羊户的投资额是决定适度规模的主要因素。根据投资和羊群基础母羊数量设计的三种典型经营条件下，基础母羊全部留种的经营方式可获最大投资收益率(140．1％)；基础母羊数量达到20只以上，自繁种公羊的经营方式的最大投资收益率为74．0％；资金有限，人工授精配种的经营方式的投资收益率最低，为73．1％。     

维生素E对绵羊鲜精及冻精精液品质的影响

日粮中添加维生素E可以提高绵羊鲜精的活率 ,对照组和试验组的活率分别为 0 72± 0 0 7和 0 78± 0 0 6(P <0 0 5 ) ,显著改善鲜精精液品质。采用两步稀释法在绵羊冷冻精液稀释液中添加维生素E ,可以极显著降低冷冻对精子顶体的冷刺激损害程度 ,试验组的活率 ( 0 43 5± 0 0 2 0 )极显著高于对照组 ( 0 3 65± 0 0 2 6) (P <0 0 1) ,精子顶体的总异常率试验组 ( 3 3 72 % )极显著低于对照组 ( 4 4 3 5 % ) (P <0 0 1) ,其中试验组顶体膨胀率和脱落率与对照组分别为 1 2 8%±0 5 8%、2 3 0 8%± 1 45 %与 2 68%± 0 5 9%、2 8 96%± 3 14 %。冷冻精液品质显著改善。     

丝状霉形体丝状亚种SC型脂蛋白LppQ N末端基因克隆与序列分析

脂蛋白LPPQ是丝状霉形体丝状亚种SC型(MmmSC)非洲株、欧洲株和疫苗株所特有的。LPPQ N末端域具有良好的免疫原性，在牛体内可诱导产生强大、特异、早期、持续的免疫反应。本研究根据已发表的LPPQ基因序列设计引物，用Pyrobest^TM高保真DNA聚合酶从MmmSC HVRI X株中扩增出了LPPQ N末端基因序列，并进行了克隆与序列测定。核苷酸序列比较结果显示，HVRI X株的LPPQ N末端基因序列与国外发表的序列同源性为99．7％，由其推导的氨基酸序列同源性为99，1％，为脂蛋白LPPQ N末端基因体外表达奠定了基础。     

仔猪小肠肌间神经丛NDP阳性神经元形态的定量研究

采用小肠铺片NADPH黄递酶组化染色和细胞影像分析的方法对0、5、28日龄的仔猪小肠肌间神经丛神经元群体的形态参数进行定量测定。结果表明:随日龄增长,小肠肌间神经元胞体面积增大,出生后的头几天尤为明显。NDP阳性肌间神经元胞核面积平均值与胞体面积平均值呈正相关。胞体面积平均值在0日龄为234.98±23.48μm2,5日龄为346.30±33.07μm2,28日龄为364.17μm2;胞核面积平均值在0日龄为69.85±6.27μm2,5日龄为88.25±2.39μm2,28日龄为84.15μm2。核质比随日龄呈下降趋势,核质比在0日龄为0.298±0.003,5日龄为0.257±0.027,28日龄为0.231。上述测量值不仅表现出日龄差异,而且在同一日龄小肠的前后段亦有所不同。NDP阳性神经元胞体和胞核大小都以十二指肠最大,空肠前段次之,回肠最小。0日龄仔猪神经元胞体面积分布在100～300μm2之间的占70.79%,5日龄和28日龄胞体面积分布在100～400μm2之间的分别占64.81%和63.22%。肌间神经丛NDP阳性神经元主要为Dogiel 型神经元。