Selection and amplification of the liver stem cell subset from rat bone marrow cells with a medium containing cholestatic serum in vitro

AIM: To explore the feasibility of direct separat and selective enlargement of the bone marrow-derived liver stem cells (BDLSC) from bone marrow cells with a culture system containing cholestatic serum in vitro. METHODS: Bone marrow cells of rats were cultured with selective media containing 2%, 5%, 7% and 10% cholestatic rat serum, respectively. The BDLSC were then induced to proliferate with the addition of hepatocyte growth factor (HGF) on the firth day. BDLSC were characterized using immunocytochemistry and RT-PCR for lineage markers, glycogen staining and urea synthetic assay for functions 2 weeks later. RESULTS: Bone marrow cells were unble to form colony in the presence of 2% cholestatic serum and apopotosis appeared gradually in 7% or 10% cholestatic serum. The BDLSC survived in the medium containing 5% cholestatic serum while the other types of cells did not. The survival cells proliferated with a high speed during the second week and then formed hepatocyte-like colony-forming units (H-CFU). Cells in the H-CFU expressed the characteristic proteins of fetal hepatocytes. Furthermore, they had glycogen storage and urea synthesis functions, two of the critical features of hepatocytes. CONCLUSION: The selective micro-environment effectively selected BDLSC from the bone marrow cell, and will be a new way to provide an abundant source of donor hepatocytes for clinical cell therapy.     

离体条件下水稻种衣剂对串珠镰刀菌生长及形态毒理影响

 检测了黑龙江水稻主产区4个当地主栽品种的种子内部镰刀菌寄藏情况,测定了20%克福甲和20%克多甲种衣剂对种子带菌消毒处理效果及对水稻串珠镰刀菌(Fusarium moniliforme)的抑菌作用和联合毒力,并借助扫描电镜观察了上述2种混配种衣剂对串珠镰刀菌的形态毒理影响。结果表明,种子内部镰刀菌的分离频率高达56.7%~96.0%,其中串珠镰刀菌的分离频率为32.6%~48.2%,2种种衣剂对带菌种子具有显著的消毒处理效果。20%克福甲和20%克多甲种衣剂对镰刀菌F.moniliforme的毒力指数分别为457.11和802.04,增效倍数分别为6.53和0.13。20%克多甲种衣剂(多菌灵:甲基立枯磷为5:5,W/W)对串珠镰刀菌的抑菌作用优于20%克福甲种衣剂(福美双:甲基立枯磷为8:6,W/W),增效作用低于20%克福甲种衣剂。电镜观察表明,种衣剂低浓度至高浓度处理下均可引起串珠镰刀菌菌丝不同程度的异常生长,表现为主菌丝局部膨大或形成菌丝束,菌丝顶端异常膨大、缢缩或形成穗状和花絮状分枝。     

引起糖甜菜细菌性叶斑病的萎蔫短小杆菌新致病变种

 1995年在内蒙古临河市新发现了糖甜菜细菌性叶斑病,从病斑所分离的10个细菌菌株经柯赫氏法则验证,均确系该病的病原菌。采用形态观察、表型特征和生理生化特性测定、数值分析、血清学反应、细胞化学成分分析、DNA G+C mol%和DNA-DNA同源性测定进行了鉴定,并与植物病原棒形细菌15个标准菌株进行了比较。该病原菌为革兰氏阳性细菌,不规则短杆状,有一根鞭毛、亚极生或侧生,结合其生理生化特性、细胞化学成分和DNA G+C mol%和DNA-DNA同源性测定结果,认为应属于短小杆菌属(Curtobacterium)的萎蔫短小杆菌(Cur. flaccumfaciens),数值分析也支持这一结论。此外,据血清学反应结果及其对短小杆菌属的其它植物寄主的致病情况,认为该病原菌应是萎蔫短小杆菌种下的一个新的致病变种,定名为Curtobacterium flaccumfaciens pv. beticola pv. nov. Chen et al.,2000(萎蔫短小杆菌糖甜菜致病变种)。     

大米粉的猪回肠氨基酸真消化率测定

大米加工副产品大米粉长期以来被添加到米糠中利用，显著降低了其利用率。本试验用三头施以回直肠吻合术猪，采用单因子设计，以大米粉为半纯合日粮，无氮日粮由醋酸纤维素提供粗纤维来测定内源值，测定了大米粉的猪回肠末端氨基酸表观消化率和真消化率。大米粉的回肠末端氨基酸的表观消化率的平均值为76．20％，真消化率的平均值为85.86％。这些数据可为我国氨基酸消化率数据库提供参考，并且可作为以可消化氨基酸为基础配制日粮时的依据。     

关于修订桃种质资源(Prunus persica)描述体系的建议

评价是种质资源工作的重要环节之一,是资源利用与共享的基础。目前,中国桃种质资源的评价描述体系一般是按照中国1990年编制的“桃种质描述符”进行。通过对国际植物遗传资源委员会1980年编制“桃描述系统″和中国“桃种质描述符”在评价的层次、评价内容、评价方法和评价标准的系统比较,以及2个标准在种质资源科研实践使用过程中存在的不足之处,结合中国桃种质资源工作的特点和国外资源的描述方法的新进展,提出中国桃种质资源描述系统的修改建议,为建立中国标准化、数字化的桃种质资源描述系统奠定基础。     

Upregulation of neurokinin A levels by nerve growth factor in the experimental asthmatic guinea pig

AIM: To explore the regulatory mechanism of nerve growth factor (NGF) on neurokinin A in the experimental asthmatic guinea pig. METHODS: Radioimmunoassay was used to determine the alteration of neurokinin A levels in the lower respiratory tract and visceral sensory afferent sites while NGF was absent (inhalation of NGF antibody through nasal cavity) in the asthmatic guinea pig. RESULTS: The contents of neurokinin A in the trachea, bronchus, lung, C₇-T₅ spinal ganglia and the correspondent spinal dorsal horn, nodose ganglia and solitary nucleus area in the experimental asthmatic guinea pig with the absent of NGF in the respiratory tract were much lower than those in the asthmatic and control groups (P＜0.01). CONCLUSION: NGF upregulated the contents of neurokinin A in the lower respiratory tract and visceral sensory sites of the experimental asthmatic guinea pig, and both might be involved in the pathogenesis of asthma.     

Bone marrow mesenchymal stem cells are induced to differentiate into cardiomyocytes by hepatocyte growth factor in vitro

AIM: To explore a new method of hepatocyte growth factor (HGF) inducing bone marrow mesenchymal stem cells (MSC) to differentiate into cardiomyocytes. METHODS: Bone marrow MSC was cultured with DMEM media (10% fetal calf serum) 4-6 passages, and induced by HGF (10 μg/L) for 30 d. Automatical beating of the differentiated cells was observed daily with transverse microscopy, or under condition of 0.1% isoproterenol or cal-cium-deprived incubation. Specific cardiac myosin in the cells was indentified by immunochemistry. RESULTS: At 14-20 d of differentiation, bone marrow mesenchymal stem cells formed clones, in 10%-50% of which spontaneous beating cell-mass had come to continuously exist. Isoproterenol increased the beating rate and calcium-deprived media inhibited the beating. The cells were identified to be cardiomyocytes by expression of cardiac myosin heavy chain. CONCLUSION: HGF may induce bone marrow mesenchymal stem cells into cardiomyocytes with high efficiency, but the differentiating pathway of stem cells remains to be further studied.     

锌对雏鸭外周血T-淋巴细胞的影响

15 0只 1日龄天府肉鸭健雏随机分为 3组 ,分别喂给缺锌 (每千克日粮含 Zn 2 2 .9mg)、对照 (每千克日粮含 Zn10 0 mg)和锌中毒 (每 kg日粮含 Zn130 0 mg)日粮 7周或 4周 ,以酸性 - α-醋酸萘酯酶 (ANAE)染色法观测外周血 T-淋巴细胞的动态变化。结果锌缺乏组和锌中毒组雏鸭外周血 T-淋巴细胞的 ANAE阳性率显著低于对照组 (P<0 .0 1) ,表明锌缺乏或锌中毒可抑制 T-淋巴细胞的生成 ,降低其在外周血中的数量。本试验还对锌缺乏和锌中毒引起的外周血 T-淋巴细胞减少的机理进行了探讨     

家鸡Leptin成熟肽cDNA的克隆、重组蛋白表达及纯化

从 18周龄鸡卵巢组织中抽提总 RNA,使用六聚体随机引物反转录后 ,用鸡 L eptin(瘦素 )特异性引物扩增出鸡L eptin编码区第 5 2～ 4 6 0 bp的长度为 4 0 9bp的 c DNA片段。根据鸡 L eptin的 3′端第 4 6 0～ 4 92 bp序列设计了 3条部分相互重叠并且顺序串联延伸的下游反义引物 ,并在最后 1条引物 3′端连接上 Eco R 切点 ;在以上用于反转录扩增的 5′端引物的 5′端连接一 Bam H 切点。用该 5′端引物分别与 3个 3′端引物配对 ,利用反转录扩增出的 4 0 9bp的L eptin c DNA片段作为第 1模板进行扩增 ,扩增产物再作为模板与下一引物对再次扩增。经 3次扩增得到编码鸡 L ep-tin成熟肽的全长 4 5 1bp的 c DNA序列。将该 4 5 1bp的 L eptin c DNA序列经 Bam H 和 Eco R 双酶切后 ,克隆入表达质粒 p RSET A的 Bam H 和 Eco R 两酶切位点之间 ,构建成表达质粒 p L ep- SCAU。转化有重组表达质粒 p L ep-SCAU的大肠杆菌 BL 2 1(DE3)在 L B培养基中培养后 ,经 IPTG诱导表达出相对分子质量为 2 0 10 0的鸡 L eptin融合蛋白和少量 4 0 2 0 0的 L eptin融合蛋白。L eptin融合蛋白的表达在 IPTG浓度为 0 .0 5 mmol/ L 时达到最高 ,占总菌体蛋白的 32 .6 %。用 Ni- NTA凝胶从 7L 发酵培养菌裂解液中纯化出 180 m g左右     

大肠杆菌新型菌毛(F1987)的基因定位

对动物产肠毒素性大肠杆菌 (ETEC)的一种新型菌毛 (F1987)进行了相应基因的定位研究 ,通过对表达该新型菌毛 (F1987)相应 ETEC菌株的培养、质粒提取、对大肠杆菌受体菌株 DH5α的转化及阳性转化子筛选与相应菌毛表达的检定等 ,初步表明该新型菌毛 (F1987)的基因定位于质粒上