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.     

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.     

酯酶同工酶在食用菌菌种选育中的应用研究

采用聚丙烯酰胺凝胶电泳 ,分析了不同培养时间和不同培养基种类对平菇、杏鲍菇、白灵菇酯酶同工酶的影响 ,结果表明在不同培养时间和不同培养基条件下 ,酯酶同工酶酶谱的酶带呈现不同的多态性 ,由此提示同工酶在食用菌菌种选育的应用上应固定最佳培养时间及最适培养基种类 ,并在所有操作条件一致的情况下 ,才能得到稳定准确的分析结果     

农民旅游市场开发浅议

近几年 ,我国国内旅游发展迅速 ,城镇居民旅游市场日益成熟 ,无论出游人口还是旅游花费都明显增加 ,但农民旅游市场却没有得到应有的重视 ,旅游供给严重不足。鉴于农民旅游的重要地位 ,本文分析了当前农民旅游特征及其发展潜力 ,在此基础上尝试性地提出了开拓农民旅游市场的几点措施建议。     

Bt玉米对中红侧沟茧蜂发育的间接影响及室内行为研究

利用Y-型嗅觉仪研究了中红侧沟茧蜂(Microplitis Mediator Haliday)对Bt玉米、常规玉米、粘虫幼虫和其虫粪及虫害苗挥发物的行为反应,同时研究了Bt玉米对中红侧沟茧蜂发育的影响。结果表明,两种玉米健康植株的挥发物对中红侧沟茧蜂均有引诱作用;中红侧沟茧蜂对两种玉米的健康植株及机械损伤株挥发物之间的选择性无显著差异;相对于Bt玉米,中红侧沟茧蜂更趋向于选择常规玉米的虫伤苗及玉米-粘虫幼虫-虫粪混合物的挥发物。与对照(寄生于取食常规玉米粘虫的中红侧沟茧蜂)相比,寄生于取食Bt玉米粘虫幼虫的中红侧沟茧蜂幼虫历期延长,出茧率、茧重、羽化率、蜂重均有显著降低,茧历期、蜂历期则差异不显著。     

Osmolarity,cell volume and proliferation in nasopharyngeal carcinoma cells

AIM: To investigate the relationship between osmolarity, cell volume and cell proliferation in nasopharyngeal carcinoma cells. METHODS: MTT method was applied to detect the proliferation ability of the poorly-differentiated nasopharyngeal carcinoma cell (CNE-2Z) under various osmolarity conditions. The flow cytometry was used to analyse cell cycle distribution. Cell volume was obtained by the image analysis of living cells and cell viability was determined by the trypan blue assay. RESULTS: Cultivation of cells under the hypertonic conditions of 370 and 440 mOsmol/L increased cell volume by 8.7% and 27.8% and facilitated cell proliferation by 22.2% and 33.9%, respectively. However, hypotonic incubation of cells with osmolarity of 160 and 230 mOsmol/L decreased cell volume by 12.8% and 4.1% and inhibited cell proliferation by 34.0% and 15.6%, respectively. Cell volume was positively correlated with cell proliferation rate. Long-term cultivation of cells under anisotonic conditions did not significantly alter cell cycle distribution, but hypotonic cultivation decreased cell viability. CONCLUSION: Proliferation of nasopharyngeal carcinoma cells was closely correlated with the osmolarity of culture medium and cell volume. Hypotonic cultivation may inhibit cell proliferation by decreasing cell volume to facilitate cell death mechanisms.     

Effect of YIGU capsule on IGF-I mRNA and protein expression in rat osteoblasts in vitro

AIM: The effects of YIGU capsule on proliferation and IGF-I mRNA protein expressions in osteoblasts were studied. METHODS: (1) Forty 12-month old Sprague-Dawley female rats were divided randomly into four groups (YIGU capsule high dose group, medium dose group and low dose group; saline group), the drug-containing serum and control serum were prepared. (2) The new-born Sprague-Dawley rat osteoblasts were cultured with different YIGU capsule drug-containing serum at different concentrations and different exposure time. MTT method was used to observe proliferation of osteoblasts. (3) RT-PCR method was used to measure the relative IGF-I mRNA levels and ELISA method was used to measure IGF-I secretion at different exposure time. (4) ELISA method was used to measure IGF-I secretion at different exposure time. RESULTS: (1) Proliferation of osteoblasts was more than the control groups after 48, 72 and 96 h, respectively (P<0.01); (2) The relative IGF-I mRNA levels and IGF-I protein expression were higher than those in control group after 48, 72 and 96 h, respectively (P<0.01 or P<0.05). CONCLUSIONS: It was suggested that YIGU capsule drug-containing serum promoted proliferation, IGF-I mRNA and protein expression. These results may be parts of the mechanisms of YIGU capsule to prevent and treat osteoporosis.     

体外培养条件对延边黄牛受精卵体外发育的影响

从延边黄牛卵巢中采集未成熟的卵母细胞进行体外成熟培养、体外受精及受精卵体外培养。结果表明 :1将卵子用 2种不同的培养液进行体外成熟培养和受精卵体外培养 ,TCM199组的卵裂率 (5 6 .3% )极显著高于 D- PBS组(33.6 % ) (P<0 .0 1) ;TCM199组的囊胚发育率和孵化率 (15 .1%、13.7% )虽高于 D- PBS组 (10 .5 %、8.7% ) ,但 2个组之间无显著差异 (P>0 .0 5 )。2以 TCM199作为基础培养液 ,分别用含激素培养液和不含激素培养液进行成熟培养和受精卵体外培养 ,添加激素组的卵裂率、囊胚发育率及囊胚孵化率 (81.2 %、17.5 %、15 .3% )高于没有添加激素的对照组 (75 .8%、12 .1%、10 .5 % ) ,但 2个组之间无显著差异 (P>0 .0 5 )。 3体外受精卵与单层颗粒细胞共培养组的卵裂率、囊胚发育率及囊胚孵化率 (78.0 %、11.5 %、9.9% )显著高于非共培养组 (6 8.1%、5 .4 %、3.6 % ) (P<0 .0 5 )。     

猪卵泡卵母细胞体外成熟与冷冻保存

研究了激素、猪卵泡液、不同类型血清对猪卵泡卵母细胞体外成熟的影响 ;比较了卵泡直径小于 2 mm、2～ 5 mm和大于 5 mm的卵母细胞体外成熟能力的差异 ,并对不同发育阶段猪卵母细胞的冷冻保存进行了研究。结果表明 :猪卵母细胞体外培养 4 8h时 ,培养的前 2 4 h培养液中加入激素 ,后 2 4 h去掉激素 ,卵母细胞的 A级成熟率 (5 1.73% )和总成熟率 (83.2 5 % )最高 ,极显著高于前 2 4 h不加激素 ,后 2 4 h添加激素培养的成熟率 (P<0 .0 1) ;也显著高于不含激素的培养液连续培养 4 8h的成熟率 (P<0 .0 5 ) ;但与添加激素连续培养 4 8h的成熟率差异不显著 (P>0 .0 5 )。在体外成熟培养液中 ,添加 10 % (体积分数 ) ECS的成熟率 (72 .86 % )显著高于添加 10 % (体积分数 ) NCS的成熟率(6 2 .2 1% ) (P<0 .0 5 ) ,而添加 10 % p FF则抑制卵母细胞的体外成熟。随着卵泡直径的增大 ,卵母细胞体外成熟能力逐渐增强。采用程序冷冻保存方法 ,成熟卵母细胞的成活率 (35 .5 9% )显著高于培养前 (2 4 .6 4 % )和培养 2 4 h(2 3.36 % ) (P<0 .0 5 )。培养 2 4 h(77.2 2 % )和培养成熟 (72 .81% )的卵母细胞解冻后的形态完整率均显著高于培养前(5 3.2 4 % ) (P<0 .0 5 )