The application of atomic force microscope in clinic diagnosis of erythrocytes

AIM: To investigate the application of atomic force microscope (AFM) in clinic diagnosis. METHODS: Topographic images and some parameters of large range field and microstructures of erythrocytes in the blood of normal subjects, lung cancer and myelodisplastic syndrome (MDS) patients were examined by atomic force microscope. RESULTS: Many clear topographic images of many erythrocytes, single erythrocyte, and microstructure of erythrocyte membrane surface were obtained. Many erythrocytes in lung cancer patients were found to change into echinocytes. One erythrocyte had 10-20 protuberances, most of which, with a mean width of 589. 0 nm and a length of 646. 7 nm, were on the edge of cells. The protuberances on the center of echinocytes are lodged and embedded. The erythrocytes of MDS patients were biconcave in shape. Many apertures with different diameters of tens to hundreds nanometer appeared on the surface of cell membrane. CONCLUSIONS: AFM can be widely applied in clinic pathological inspection, including quantification of cells, obtainment and comparison of many parameters (such as diameter, thickness, volume, surface, surface area/volume ratio), observation of topograph of single cell, and observation and comparison of membrane surface microstructure of cells, and so on.     

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.     

Study on adherent precursors from human cord blood

AIM: To confirm the existence of the endothelial progenitor cells in human cord blood and to study its differentiation and development process. METHODS: The mononuclear cells in human cord blood were isolated using lymphocyte separation solution. Then the mononuclear cells were cltured in MCDB131 containing 20% fetal bovine serum. The effects of 5 μmol/L dexamethasone, the extract from bovine brain, insulin and hypoxanine on the proliferation and differentiation of the adherent cells were observed. The morphology of the adherent cells were examined twice daily by inverted phase contrast microscope. CD34 and CD14 expression were determined by FACS. Immunohistochemistry was used to confirm the expression of factor Ⅷ. RESULTS: The proliferative endothelial progenitor cells existed within the CD34^-adherent mononuclear cells of human cord blood. Dexamethasone and hypoxanine decreased the number of spindle-shaped cells and caudated cells. Bovine brain extract, insulin and FCS enhanced the number of spindle-shaped cells and caudated cells. CONCLUSION: The existence of endothelial progenitor cells within the CD34 - adherent monouclear cells of the human cord blood was observed and these cells were able to differentiate into endothelial-like cells in vitro.     

腐败组织中毒鼠磷的GC／NPD、GC／ECD检验

本文初步建立了一套混合溶剂提取、中性层析氧化铝小柱净化及GC／ECD和GC／NPD分析的腐败生物样材中毒鼠磷的检验方法。GC／ECD的检出限为1ng;GC/NPD的检出限为20ng。     

粉蕉、大蕉和龙牙蕉的AFLP分类研究

 应用AFLP 技术, 对32 个粉蕉、大蕉、龙牙蕉品种进行了遗传多样性分析及分类研究。在分子水平上, 可将供试品种聚为6 个独立群体, 其中包括3 个分别由单一品种组成的群体(群体Ⅳ: 酸大蕉;群体Ⅴ: 孟变; 群体Ⅵ: 千旁培) ; 可以将群体Ⅰ (大蕉) 和群体Ⅱ (粉蕉) 分别划分为3 个品种群, 将群体Ⅲ (龙牙蕉) 划分为2个品种群。参照Simmonds 的形态学分类方法, 研究结果显示粉蕉、大蕉以及龙牙蕉各个品种之间存在丰富的遗传多样性, 将其划分为AAB 或ABB 等类型, 不能反映品种间的亲缘关系; 粉蕉与大蕉两个类群之间遗传距离差异明显, 将其简单的归为ABB 类型是不妥当的。     

灰树花不同菌株液体深层培养蛋白质含量的测定

在液体培养条件下测定了发酵液中5个灰树花菌株蛋白质含量的变化。结果表明，接种后第6天发酵液中蛋白质含量最高，Gf961菌株发酵液中的蛋白质含量为0.58%，Gf962菌株发酵液中的蛋白质含量最低，为0.30%，随着培养时间的延长，发酵液中蛋白质含量逐渐下降，接种后第9天最低，随后以呈上升趋势。但上升的幅度较小。试验为确定从发酵液中提取蛋白质的最佳时期提供了一定的理论依据。     

苹果粗皮病与锰含量的关系

对胶东半岛部分发生苹果粗皮病(IBN)的果园的土壤和植株进行了分析测定,结果表明:发生苹果粗皮病的果园土壤有效锰含量较高,而且粗皮病严重果园高于粗皮病较轻果园。发生粗皮病的苹果树花器官中锰含量较高,粗皮病最严重的姜格庄果园苹果树花器官中锰平均含量比观水镇高出139.28mg/kg,比季家埠重病园和轻病园分别高122.34和182.19mg/kg。从4月到10月,苹果粗皮病逐渐加重,叶片中锰含量逐步增高。10月份姜格庄果园最高锰含量达1717.85mg/kg,平均锰含量达1119.69mg/kg,而粗皮病较轻的季家埠果园则仅306.03mg/kg,前者比后者高265.88%。对不同部位锰含量测定表明,锰含量在叶片中最高,其次是韧皮部,木质部最低。表明胶东半岛苹果粗皮病的发生是因为锰过量。可能是土壤锰含量高,pH值低,降雨多,施肥不合理等,造成土壤锰有效性增加.导致树体吸收过量而出现粗皮病害。     

桃流胶病研究进展

流胶病为桃树生产上的主要病害之一,发生普遍,危害严重。综述了引起桃流胶病的致病菌种类,病原菌的发生与侵染规律,组织病理学研究,田间自然条件与人工接种的抗性鉴定及筛选出的抗性品种,所采取的综合防治措施,以及在抗性育种与抗病性筛选的生理生化指标方面的研究进展,并对今后的工作重点提出了建议。     

Inhibitory effect of berberine on the activation and proliferation of T lymphocytes

AIM: To investigate the effect of berberine (Ber) on the activation and proliferation of T lymphocytes and its mechanism of action. METHODS: Whole peripheral blood from normal subjects was stimulated with phytohemagglutinin (PHA) or phorbol ester (PDB) plus ionomycin (Ion) and the expression levels of CD69 and CD25 were evaluated with flow cytometry after the staining with appropriate fluorescent monoclonal antibody. The distribution of cell cycles was analyzed by propidium iodide staining and dead cells by 7-aminoactinomycin live staining. RESULTS: 100 μmol/L and 50 μmol/L of Ber had significant inhibition of the expression of CD69 on T cells stimulated with PDB plus Ion or PHA, while effect of 25 μmol/L Ber was not significant. And as time of action extended, the extent of inhibition decreased. For the expression of CD25, Ber at the concentrations as above all exerted significant inhibitory effect in a dose-dependent manner. Moreover, Ber could block lymphocytes cell cycle progression from G₀/G₁ phase to S and G₂/M phase without phase specificity. Besides, live staining analysis revealed that Ber did not have significant cytotoxicity on lymphocytes. CONCLUSIONS: Ber significantly inhibits the expression of early and mid activation antigens of T cells and also blocks the progression of lymphocytes cell cycles. These results suggest that Ber exerts immunosuppression effect through inhibiting the activation and proliferation of T cells.     

复合微生物肥料在无公害蔬菜栽培上的效应初报

施用复合微生物肥料使番茄产量提高 7.3 %～ 11.1% ,可溶性固形物含量提高 0 .9%～ 1.3 % ,硝酸盐含量降低 2 7.2 %～ 3 4 .8% ;苦瓜产量提高 10 .7%～ 16.1% ,硝酸盐含量降低 2 8.7%～ 3 6.1% ;菜薹产量提高 7.2 %～ 15.1% ,硝酸盐含量降低 55.6%～ 61.5%。各处理产品中的硝酸盐含量及有害元素的含量 ,均符合无公害蔬菜产品质量标准要求