Metallothionein inhibits homocysteine-induced proliferation of rat vascular smooth muscle cells

AIM:To investigate the effect of metallothionein(MT) on proliferation of rat vascular smooth muscle cells (VSMCs) stimulated by homocysteine and its mechanism. METHODS:VSMCs proliferation was measured by [³-H]-TdR incorporation, mitogen-activated protein kinase(MAPK)activity were determined by immunoprecipitation method, the intracellular contents of MT and malondialdehyde (MDA)were assayed by -hemoglobin saturation method and TBA reaction, respectively, and lactate dehydrogenase (LDH) leakage was measured by NADH oxidation. RESULTS:Hcy(10^-6-10^-4 mmol/L) stimulated [³-H]-TdR incorporation by the VSMCs in a concentration-dependent manner. Compared with control, [³-H]-TdR incorporation in VSMCs treated with 0.1 mmol/L Hcy was increased by 4.2 fold (P＜0.01). Meanwhile, Hcy enhanced MAPK activity, MDA formation and LDH release (P＜0.01)in a concentration-dependent manner. Treatment of VSMCs with MT alone did not change above parameters, compared with control. However, MT (10^-6-10^-4 mol/L)attenuated significantly Hcy-stimulated proliferation of VSMCs (P＜0.01)in a concentration-dependent manner. And MT inhibited obviously Hcy-induced activation of MAPK activity, MDA formation and LDH release. Preincubation of VSMCs with 0.5 mmol/L ZnCl₂ for 6 h induced an increase cellular MT content by 5.7-fold (P＜0.01). The MT-overexpressed VSMCs resisted Hcy-stimulating action on MAPK activity, MDA formation and LDH leakage (P＜0.01). CONCLUSION:These results show that MT has an inhibitory effect on Hcy-induced VSMCs proliferation, and that MT could inhibit Hcy-stimulated MAPK activity and lipid peroxidation.     

In vitro growth inhibition effects of rhHGF/cHGF on SMMC-7721 human HCC cell line

AIM:To examine the effects of recombinant human hepatocyte growth factor(rhHGF) and native calf HGF(cHGF) on SMMC-7721 human hepatocellular carcinoma(HCC) cell line. METHODS:Human HCC cell line culture, photometric assay, and flow cytometric assay were used in this study .RESULTS:A similar type of dose-dependent cell growth inhibition effect on SMMC-7721 human HCC cells by rhHGF(5-20 μg/L) as well as by cHGF(25-100 mg/L) had been found, with the maximal effect at the highest concentration used. Approximately over 50% of the cells treated with rhHGF(5 μg/L, 10 μg/L, 20 μg/L) accumulated in the quiescent G₀/G₁ phase of the cell cycle over incubation periods for 3 d. CONCLUSION:The growth of SMMC-7721 human HCC cells was strongly inhibited by both rhHGF and cHGF. This might be because the cells exposed to HGF became arrested in the G₀/G₁ phase.     

Establishment of chronic obstructive pulmonary disease rat models by passive cigarette smoking and intratracheal instillation of lipopolysaccharide

AIM:To establish rat chronic obstructive pulmonary disease(COPD) models by passive cigarette smoking plus intratracheal instillation of lipopolysacchride(LPS) or passive cigarette smoking only, which would be similar to the pathogenesis of human COPD. METHODS:48 Wistar rats were randomly divided into 4 groups.(1) Healthy control I group(n=12), rats were bred 4 weeks;healthy control II group(n=12), rats were bred for 3months. (2) Model group I (n=12), 200μg lipopolysaccharide(LPS) was instilled intratracheally once for every two weeks and the rats were exposured to 5% of cigarette smoke, 0.5 h/d for 4 weeks.(3) Model group II(n=12),rats were exposed to 5% of cigarette smoke, 0.5 h/d for 3 months. The pathologic changes of airways and lung tissues, pulmonary function and blood gas analysis were determined. The airway wall lymphocytes and alveolar macrophages were counted. The cross areas of epithelial layer, smooth muscle layer and lamina propria of bronchi were measured. The hydroxyproline of lung tissue homogenates was determined by biochemistry method.RESULTS:The pathologic changes of airways and lung tissue of two models were similar to but milder than those of COPD patients(biopsy data). The collagen deposition and the cross areas of epithelial layer and smooth muscle layer in airway walls of two model groups were significantly increased than those of control groups(P＜0.01,P＜0.05).FEV_0.3/FVC% of two model groups, PaO₂ and SaO₂ of model I group were significantly decreased, while R_i and R_e in model I group were significantly increased than that of control I group(P＜0.05). The PaCO₂ and the counts of lymphocytes and alveolar macrophages of both model groups were significantly increased than those of the control groups (P＜0.01). Lots of alveolar macrophages had phagocyted smoke granules. The amounts of hydroxyproline of two model groups were significantly increased than those of control group((P＜0.05) and were negatively related to the FEV_0.3/FVC%, respectively (P＜0.01,P＜0.01) and positively related to airway resistance of model I group(P＜0.01). CONCLUSIONS:COPD rat models were successfully established by passive cigarette smoking plus intratracheal instillation of LPS and cigarette smoking only. The pathologic changes were similar but milder than those of COPD patients. The airway obstruction of model I group was more severe than that of model II group, but they have no significant difference.     

Effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts

AIM: To investigate the effects of PD98059 on the differentiation from mesenchymal stem cells to osteoblasts.METHODS: hMSC were separated from human marrow and expanded in cuture medium. hMSC were induced with dexamethasone, β-glycerophosphate, vitamin C which acted as osteoblast differentiation inducer. PD98059 was added into the osteoblasts induction medium. The cells were assayed with cell morphology, alkaline phosphatase (AP) activity and calcium deposition. RESULTS: The isolated cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. After induced with osteoblasts induction medium, the cells showed changes in cell morphology from spindle-shape to cuboidal and polygonal. The AP activity increased gradually and reached the peak in 12 days, then decreased. Many scattered tangerice calcium nodes were observed. PD 98059 significantly inhibited AP activity and calcium deposition in a dose-dependent manner. A striking observation of the present study was that a few adipocytes appeared in cultures that were treated with PD 98059 and osteogenic differentiation medium. CONCLUSION: These results indicated that osteogenic diferentiation from the hMSCs was related to the activation of the ERK.     

Effects of carvedilol,cilazapril and their combination on left ventricular remodeling after acute myocardial infarction in rats

AIM:To compare the effects of carvedilol, cilazapril and their combination on left ventricular remodeling(LVRM) after acute myocardial infarction(AMI) in rats. METHODS: Twenty-four hours after AMI operation, 100 surviving rats were randomly assigned to: ①AMI control(n= 25), ②AMI+carvedilol(1 mg·kg^-1 ·d^-1, n= 25)(C1), ③AMI+cilazapril(1 mg·kg^-1 ·d^-1, n= 25)(Z1), and ④ AMI+combination(n= 25) groups. Sham-operated group(n= 17) were selected randomly. After 4 weeks of therapy with the drugs gastric gavage, hemodynamic and pathological studies were performed. RESULTS: There were no significant differences in MI size among the four AMI groups(all P> 0.05) Left ventricular(LV) end diastolic pressure(LVEDP), volume(LVV), weight(LVW) and septal thickness(STh) were all higher and left ventricular pressure maximal rate of rise and fall(±d p /d t) were lower(all P< 0.01) in AMI group than sham-operated group. The LVEDP, LVV, LVW and STh were all lower and ±dp /dt were higher in Z1, C1, and combination groups than those in AMI group(P< 0.05, P< 0.01), with LVEDP and STh were more lower in the combination group than in the two monotherapy group(P< 0.05, P< 0.01), but there were no significant differences in other variables among the three therapy groups. CONCLUSION: Carvedilol, cilazapril and their combination all can prevent from LVRM after AMI in rats, improve hemodynamics and LV function, with the combination superior.     

蔗渣滤泥培养料栽培平菇试验

以不同比例滤泥代替栽培料中的麦麸进行平菇栽培试验。结果表明 ,滤泥能替代栽培料中的麦麸。     

落叶果树氮素营养研究进展

氮素作为养分与信号对果树的生长发育及产量品质的形成有重要影响。近年来有关果树的施氮效应、需氮特性、诊断技术、精准施氮技术的研究取得了许多进展,在保证果树优质丰产的前提下,为降低氮肥施用量,提高氮肥利用率,减少环境污染提供了施氮的原理与技术。     

稻瘟菌诱导性稻叶脂氧合酶的进一步纯化及抗体制备

 采用DEAE-Toyopearl离子交换、Buty l-Toyopear l疏水层析、CM-Toyopearl离子交换、Sephacryl S100凝胶过滤和FPLCMono Q等步骤,从非亲和性稻瘟菌侵染稻叶中纯化了两种诱导性脂氧合酶,即CM-Loxl和CM-Lox2。SDS-PAGE检测结果表明:CM-Lox1和CM-Lox2为单链多肽,它们的分子量分别为98 kd和102 kd。利用微量免疫法制备了CM-Lox1和CM-Lox2的抗体,制备的抗体效价达到1:1000倍,能检测0.5~2.0 ng的抗原。免疫印迹杂交证明CM-Lox1和CM-Lox2之间存在密切的血清学关系。此外,Anti-CM-Lox1和Anti-CM-Lox2与水稻中另一稻瘟菌诱导性脂氧合酶RLL以及发芽种子出现的RL-2也存在交叉反应。     

江汉平原水稻白叶枯病菌致病型的变异研究

 从水稻和稻田周围常见的10种杂草上来集梨孢属菌株,以涂沫法作交互接种,分离自牛筋草、狗尾草、法氏狗尾草、李氏禾、罗氏草、铺地黍、稗和马唐的梨孢菌与分离自水稻的梨孢菌可以交叉侵染其寄主,茭白和碎米莎草梨孢菌与稻梨孢菌不能交互侵染;自然诱发条件下来自牛筋草和狗尾草的梨孢菌与来自水稻的梨孢菌对其寄主同样发生交叉侵染。直接从杂草寄主上分离的梨孢菌株对水稻的致病性较弱,接种到水稻感病品种发病后再分离的菌株,对水稻致病性增强,对原寄主的致病性基本保持不变。推断稻瘟病区稻田周围牛筋草、狗尾草、法氏狗尾草、李氏禾、罗氏草、铺地黍、稗和马唐等杂草寄主上的梨孢菌在稻瘟病菌的积累和传播方面具有作用。     

玉米螟赤眼蜂适宜生境的研究与利用:Ⅲ.夏玉米间作匍匐型绿豆对赤眼蜂的增诱作用及其在穗期玉米螟防治中的利用

本文就夏玉米间作匍匐型绿豆对玉米螟赤眼蜂增诱作用及其在穗期玉米螟防治中利用的可行性进行了研究。结果表明，夏玉米间作匍匐型绿豆结合接种式放蜂区，与平作夏玉米结合接种式放蜂区和不放蜂对照区相比，明显提高了玉米螟赤眼蜂对螟卵的寄生率，１９９２年三者人工挂卵的卵块总寄生率分别为２６．７％、３．６％和０；１９９３年分别为２８．０％、６．６％和１．０％，自然落卵的寄生率分别为６９．１％、２２．４％和４．６％。１９９４年用心叶期抗螟品种间作绿豆结合接种式放蜂区，人工挂卵的卵块总寄生率为３０．７％，而平作夏玉米不放蜂对照区仅为２．５％；自然落卵的卵块寄生率前者为５６．２％，后者为１２．３％。心叶期抗螟品种间作匍匐型绿豆结合接种式放蜂区、心叶期抗螟夏玉米平作不放蜂和感螟品种不放蜂对照区的平均百株蛀孔数分别为５４．８、１０２．６和２７７．２个，放蜂区与两个不放蜂对照区相比，对玉米螟的防效分别为４６．６％和８０．２％。试验结果显示采用心叶期玉米抗螟品种间作匍匐型绿豆，并接种释放少量赤眼蜂等综合措施对穗期玉米螟害有明显的控制作用