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。     

Epoxyeicosatrienoic acid: the signal transduction

Cytochrome P450 monoxygenase converts arachionic acid to four epoxyeicosatrienoic acid regiosomes: 5, 6-EET (epoxyeicosatrienoic acid); 8, 9-EET; 11, 12-EET and 14, 15-EET. Recent studies show that EETs are involved in signal transduction. EETs open Ca²⁺-sensit ive K⁺ channel and inhibit Na⁺ channel, Ca²⁺-sensitive Cl^- channel and so on. What is more, EETs have been demonstrated to activate PP60^c-src and initiate a tyrosine kinase cascade that mediates mitogenic effects.     

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.     

Study on the electrophysiological characteristics of ion channels of stem cell derived cardiomyocytes of mouse

AIM: To study the electrophysiological characteristics of ion channels of stem cell derived cardiomyocytes(SCDC) of mouse. METHODS: Embryonic stem cells of D₃ line(ES-D₃) were cultured on the MEF feeder layer with BRL conditioned medium, and fetal mouse heart cells(FMHC)were cultured in vitro. Then ES-D₃ cells were induced to differentiate into many kinds of cells. SCDC were harvested on day 12 after differentiation initiating and identified by electro-microscope and immunocytochemistry. SCDC and FMHC were prepared for the patch-clamp research. Sodium and calcium currents together were elicited and compared between SCDC and FMHC. RESULTS: The current characteristics of sodium and calcium channels of SCDC were very similar to FMHC. CONCLUSION: The functional expression of ion channels occurred during ES-D₃ cells differentiation and the electrophysiological characteristics of sodium and calcium channels of SCDC are very similar to FMHC.     

Study on integrin-mediated albumin microbubbles adherence to activated leukocytes

AIM:To investigate the mechanism responsible for albumin microbubbles adherence to activated leukocytes. METHODS: In vitro studies were performed in which activated or nonactivated leukocytes were incubated with albumin microbubbles and observed under microscopy. The suspensions of leukocytes and microbubbles which contained or absented of integrins were analyzed with flow cytometry.RESULTS: A minimum of 50cells were identified under transillumination. 5 min after microbubbles were incubated with leukocytes, the number of cells interacting with microbubbles was greater for activated cells than for nonactivated cells(20.30±2.67 vs 4.50±1.43, P <0.01).Microbubbles attached to the surface of activated leukocytes were phagocytosed and remained intact for up to 30min. Microbubble attachment was inhibited notably by blocking the leukocyte β₂-integrin Mac-1(P <0.01) and by VLA-4mAb slightly(P <0.05) CONCLUSION: The mechanism of albumin microbubbles attaching to and phagocytosed by leukocytes was due to β₂-integrin and VLA-4 mediation. Phagocytosed microbubbles can remain at the regions of inflammation for15 min, also responsible to ultrasound.     

达县分葱田甜菜夜蛾发生规律初步研究

采用系统调查方法研究了达县地区分葱田甜菜夜蛾的生活史、发生与环境的关系。甜菜夜蛾在达县分葱田1a发生6代,以三至五代为害最重,世代重叠。其数量消长与虫源、夏秋季气候、天敌、栽培管理等因素密切相关。通过调查分析提出本县初始虫源非本地虫源。     

两种高水溶性β -环糊精衍生物对甲基对硫磷的增溶和光催化降解作用

研究了两种β-环糊精的衍生物甲基-β-环糊精(MCD)和羟丙基-β-环糊精(HPCD)对甲基对硫磷的增溶作用和对紫外光降解的影响。结果表明:MCD和HPCD能增强甲基对硫磷的水溶性,在25℃下,20 g/L的MCD和HPCD溶液中,甲基对硫磷溶解度比在纯水中分别提高了21.91和17.92倍;另外,3 g/L、6 g/L MCD和HPCD分别处理的甲基对硫磷,其光降解速率分别加快了4.87~6.85倍。增溶作用和光敏效应主要是由于MCD和HPCD与甲基对硫磷形成包合物引起的。     

国产克洛素隆尿药排泄数据分析

应用Kromasil-C18色谱柱(250 mm×4.6 mm,5 μm),WatersTM480型可调波长紫外检测器,0.01M磷酸钾(pH=7):乙腈(3:1)为流动相,检测波长265 nm,含量测定采用标准曲线法,建立了RP-HPLC法检测绵羊尿中克洛素隆含量的方法.方法有效性评价结果表明,尿药含量在0.01～5.0μg/ml及5.0～30.0μg/ml范围呈良好线形关系(r=0.9993、0.9995),方法平均回收率99.32%,日内、日间变异系数分别为3.91%、6.28%.尿药最低检测限0.005μg/ml.试验绵羊以7 mg/kg单剂量经静脉、肌肉及口服三种途径给药后,尿中药物浓度分析表明,克洛素隆经体内处理后主要经肾脏排泄,给药96 h内,静注经尿排泄原药为81.76%,肌注为64.03%,口服为48.50%.     

Effects of 11, 12-EET and ischemic preconditioning on phosphorylated ERK and p38 MAPK during ischemia and reperfusion in rat myocardium

AIM: In order to study the relationship between the ERK and p38 MAPK activation and the protection of 11, 12-epoxyeicosatrienoic acid (11, 12-EET) and ischemia preconditioning (IP), the effects of 11, 12-EET and ischemic preconditioning on phosphorylated ERK and p38 MAPK during ischemia and reperfusion in rat myocardium were examined. METHODS: The rat heart was subjected to ischemia for 5 min by ligating the left anterior descending coronary artery followed by reperfusion for 5 min (two times) to undergo ischemia preconditioning. The rats were divided into 5 groups: (1) control; (2) sham group; (3) ischemia/reperfusion (I/R) group, in which the rat heart suffered from 60 min ischemia followed by 30 min reperfusion; (4) IP plus I/R group; (5) EET plus I/R group, in which 6.28×10^-8 mol/L 11, 12-EET was injected intravenously 20 min before I/R. The heart function was examined, and phosphorylated ERK and p38 MAPK were detected by Western blot. RESULTS: At 30 min reperfusion, +dp/dt_max, -dp/dt_max and LVDP decreased significantly in I/R group compared with sham group, IP plus I/R group and EET plus I/R group; Phosphorylated ERK1/2 level was higher in I/R group than sham group, but was lower in I/R group than IP plus I/R group and EET plus I/R group; Phosphorylated p38 MAPK level was lower in control, sham, IP plus I/R and EET plus I/R group than I/R group. CONCLUSION: 11,12-EET protects rat heart against ischemia/reperfusion injury, the mechanism may be related to activation of ERK1/2 and inhibition of p38 MAPK.     

Effect of insulin on the secretion of VEGF and its receptor in serum-free cultured bovine thoracic aortic endothelial cells

AIM: This study was designed to investigate the secretion of VEGF and its receptor (flt-1 or flk-1/KDR) protein by cultured bovine thoracic aortic endothelial cells treated with various insulin concentrations. METHODS: Endothelial cells was isolated from bovine thoracic aorta, and cultured in serum-free medium, then incubated with different insulin concentrations (30 mU/L, 300 mU/L, 3 000 mU/L). The level of VEGF and its receptor (flt-1 or flk-1/KDR) protein were detected by immunohistochemical staining. RESULTS: As compared with no insulin group, the expression of VEGF protein in low insulin concentration (30 mU/L and 300 mU/L) groups were significantly increased (P＜0.01). The expression of VEGF protein in high insulin concentration (3 000 mU/L) group was significantly decreased (P＜0.05). Howerer, no difference of the expression of VEGF receptor (flt-1 or flk-1/KDR) protein among all groups (P＞0.05) was observed. CONCLUSION: Low concentration insulin up-regulates the VEGF protein expression while high concentration insulin down-regulates the VEGF protein expression in bovine thoracic aortic endothelial cells, but insulin had no directly effect on the VEGF receptor (flt-1 or flk-1/KDR) protein expression in bovine thoracic aortic endothelial cells.