Relationship between expression of MKK-4 and MMP-9 genes in primary hepatic carcinoma with or without invasion and metastasis

AIM：To investigate the expression of mitogen activated protein kinase kinase 4 (MKK-4) and MMP-9 mRNA in primary hepatic carcinoma (PHC), and to analyze its relationship with invasion and metastasis. METHODS：The expression of MKK-4 and MMP-9 mRNA in 34 cases of hepatic carcinoma tissues and adjacent tissues,and in 12 cases of normal liver tissues were detected with RT-PCR. RESULTS：(1) The expression level of MMP-9 mRNA was higher in metastatic cancer tissues than that in other tissuses (P<0.01). (2) There was significant statistical difference among the expression level of MKK-4 mRNA, but the level in metastatic cancer was low (P<0.01). (3) There was no statistical difference among the expression level of MKK-4 or MMP-9 mRNA among the adjacent tissues and normal tissues (P>0.05). (4) MMP-9 mRNA had a tendency to rise as PHC became invasive and metastatic.The expression level of MKK-4 had a tendency to decline in PHC became invasive and metastatic. (5) The expression level of MMP-9 or MKK-4 mRNA had no correlations with tumor volume,or cell differentiation (P>0.05). (6) There were correlations between expressions of MKK-4 and MMP-9 mRNA in PHC (Pearson Correlation, r=-0.925, P<0.01). CONCLUSION：There are high MMP-9 mRNA expression and low MKK-4 mRNA expression in PHC.The expression level of MKK-4 or MMP-9 mRNA is correlated with tumor metastasis.     

鸡新城疫病毒河北分离株F基因的分子特性分析

通过毒力测定、RT-PCR及F基因的序列测定与遗传进化分析,对2005-2008年从河北省部分地区的发病鸡群中分离到的10株新城疫病毒（NDV）进行了研究。各分离株经典毒力测定结果显示：MDT在37.6～54.4h之间,ICPI在1.71～2.0之间,IVPI值在2.16～2.8之间,均为新城疫病毒强毒株特征。F基因的序列测定表明,分离株之间的核苷酸序列具有77.4%～98.0%的同源性,与疫苗株Lasota的同源性为87.0%～98.9%,与国内标准强毒株F48E9同源性为89.7%～98.9%。推导其氨基酸序列分析表明,8个分离株的F蛋白的裂解位点氨基酸组成为112 R-R-Q-K-R-F117,具有NDV强毒株特征,与毒力测定结果相符,2个分离株的F蛋白的裂解位点氨基酸组成为112 G-R-Q-G-R-L117,与弱毒株特征相符。F基因分型和同源性比较显示：目前河北新城疫的流行以基因Ⅶ型为主（占70%）,同时兼有基因Ⅱ型（占20%）和基因Ⅸ型（占10%）。     

取食Cry3Aa蛋白后黄曲条跳甲成虫中肠组织的病理变化

为明确Cry3Aa蛋白对黄曲条跳甲成虫的致死效果及作用机理,采用浸叶法测定了黄曲条跳甲成虫取食Cry3Aa蛋白后的死亡情况,并利用透射电镜观察了Cry3Aa蛋白处理后成虫中肠组织的病理学变化。结果显示,取食浓度为2 mg/mL的Cry3Aa蛋白6 d和12 d后黄曲条跳甲成虫的死亡率分别为33.33%和77.78%;透射电镜观察发现,蛋白处理后成虫中肠发生了明显的病理变化,并且随着时间的延长,病理变化更加明显,主要表现为：柱状细胞变形、细胞间隙增大、微绒毛缩短并肿胀、线粒体变形、内质网肿胀。本研究表明黄曲条跳甲成虫中肠是Cry3Aa蛋白的重要作用部位,研究结果为揭示Cry3Aa蛋白对黄曲条跳甲成虫的作用机理奠定了基础。     

Effect of supplementation of blue green alga (Spirulina) on outcome of pregnancy in rats

To study the supplementary effect of Spirulina, pregnant rats were fed 5 different kinds of diets (casein, Spirulina, wheat gluten, Spirulina + wheat gluten, Spirulina-without additional vitamins and minerals), each providing 22% protein during the period of pregnancy. The outcome of pregnancy was assessed from litter and dams' weight and litter size. Maternal weight gain was found to be maximum with Spirulina + wheat gluten and least with the wheat gluten diet. Rats receiving Spirulina containing diets produced significantly (p<0.05) higher litter size than those receiving casein and wheat gluten. In spite of having higher litter size, Spirulina containing diet groups produced pups with birth weights comparable to those of casein. Spirulina appears to be a good dietary supplement during pregnancy.     

草鱼蛋白质和脂肪代谢对饥饿胁迫的响应

实验选取平均体质量(31.86±1.47) g的草鱼, 随机分为2个实验组, 对照组(CG)和饥饿组(SG), 每组3个平行, 饥饿处理15、30、45和60 d, 测定饥饿对鱼体的生长、肌肉生化组成、血液生化指标以及蛋白质代谢的影响。结果表明: 饥饿组肥满度、脏体比、肝体比显著低于对照组。随着饥饿时间的延长, 草鱼肌肉中的水分含量逐渐增加, 脂肪和蛋白质含量呈现降低趋势, 脂肪含量在4个时期没有明显的差异(P>0.05), 水分和蛋白质含量分别呈现差异性增加和降低的趋势。血清总蛋白在饥饿15 d时与对照组没有明显的差异(P>0.05), 其后3个时期对照组显著高于饥饿组(P<0.05), 但饥饿组后3个时期趋于稳定。随着饥饿时间的延长, 白蛋白、总胆固醇和甘油三脂含量在4个时期与对照组都有明显变化(P<0.05), 饥饿组呈现先降低后升高再降低的波浪形变化。丙氨酸氨基转移酶活性在15、60 d时与对照组没有明显的变化(P>0.05), 在30 d时先降低, 45 d时显著升高(P<0.05)。随着饥饿时间的延长, 与对照组相比RNA︰DNA在肌肉中没有明显的差异(P>0.05), 在肝胰脏中与对照组呈现显著性差异, 随着时间的延长饥饿组呈现先降低后增高的趋势。谷氨酸脱氢酶活性在肌肉中于饥饿15 d时与对照组没有明显差异(P<0.05), 其后比对照组显著增加, 随着时间的延长饥饿组谷氨酸脱氢酶活性呈现上升的趋势; 在15、45 d时与对照组没有明显的变化(P>0.05), 在30、45 d时显著升高(P<0.05)。肝胰脏中谷氨酸脱氢酶活性在4个时期比对照组有显著的增加; 与肌肉相比, 肝胰脏和血清中谷氨酸脱氢酶活性是一种有效的反应蛋白质代谢的指标。草鱼对体内贮能物质(蛋白质和脂肪)的动用并不是简单地与饥饿时间呈线性相关, 而是出现周期性和阶段性起伏的变化。     

Growth and Yield Response of Facultative Wheat to Winter Sowing, Freezing Sowing and Spring Sowing at Different Seeding Rates

Growth and yield of wheat are affected by environmental conditions and can be regulated by sowing time and seeding rate. In this study, three sowing times [winter sowing (first week of September), freezing sowing (last week of October) and spring sowing (last week of April)] at seven seeding rates (325, 375, 425, 475, 525, 575 and 625 seeds m^?2) were investigated during the 2002–03 and 2003–04 seasons, in Erzurum (Turkey) dryland conditions, using Kirik facultative wheat. A split‐plot design was used, with sowing times as main plots and seeding rates randomized as subplots. There was a significant year × sowing time interaction for grain yield and kernels per spike. Winter‐sown wheat produced a significantly higher leaf area index, leaf area duration, spikes per square metre, kernel weight and grain yield than freezing‐ and spring‐sown wheat. The optimum time of sowing was winter for the facultative cv. Kirik. Grain yields at freezing and spring sowing were low, which was largely the result of hastened crop development and high temperatures during and after anthesis. Increasing seeding rate up to 525 seeds m^?2 increased the spikes per square metre at harvest, resulting in increased grain yield. Seeding rate, however, was not as important as sowing time in maximizing grain yield. Changes in spikes per square metre were the major contributors to the grain‐yield differences observed among sowing times and seeding rates. Yield increases from higher seeding rates were greater at freezing and spring sowing. We recommended that a seeding rate of 525 seeds m^?2 be chosen for winter sowing, and 575 seeds m^?2 for freezing and spring sowing.     

南京典型利用方式土壤中球囊霉素含量及剖面分布特征

采用Brad-ford染色法研究了南京市5种典型利用方式土壤不同土层中(0~10、10~20、20~40 cm)球囊霉素的含量。结果表明:土壤中总球囊霉素含量为1.96~3.12 mg/g,占土壤有机碳的12.5%~29.0%,所占比例随土壤有机碳含量的增加而降低。林地和草地土壤中球囊霉素和有机碳的含量均高于3种耕作土壤(水稻田、茶园土和菜园土)。随着土层深度(0~40 cm)的增加,5种不同利用方式土壤中总球囊霉素和有机碳的含量均减小;与其他土层相比,0~10 cm土层总球囊霉素和有机碳含量均最大。耕作土壤中易提取球囊霉素更易于向总球囊霉素转化。发现5种土地利用方式下土壤中总球囊霉素含量与土壤有机碳含量极显著正相关,与土壤p H显著负相关;易提取球囊霉素与土壤有机碳含量极显著负相关。总球囊霉素和易提取球囊霉素可作为评价土壤丛枝菌根真菌活性和土壤质量的重要指标。     

水稻质体磷酸盐转运体OsPPT家族成员的功能分析

磷酸烯醇式丙酮酸/磷酸盐转运体(PPT)是植物质体磷酸盐转运蛋白家族(pPTs)成员之一,介导细胞质中的磷酸烯醇式丙酮酸(PEP)进入质体基质的同时,将磷交换到细胞质中。为对水稻OsPPT基因家族进行综合分析,探索其在水稻中的潜在功能。利用水稻原生质体瞬时转化分析OsPPT的亚细胞定位,通过酵母异源表达实验分析OsPPT的磷酸盐转运能力。设置正常供磷和缺磷等非生物胁迫水培实验处理,阐明OsPPT家族成员的组织特异性表达模式,以及对非生物胁迫逆境的响应。结果表明,OsPPT基因家族4个成员均定位于叶绿体膜,而且OsPPT可以在酵母中介导磷酸盐的跨膜转运。此外,通过实时荧光定量PCR(qRT-PCR)展示了OsPPT基因家族在应对环境胁迫时表达模式上的动态变化,比如磷饥饿,以及脱落酸(abscisic acid, ABA)、水杨酸(salicylic acid, SA)、氯化钠等非生物胁迫环境。OsPPT基因家族可能参与磷酸盐在细胞质和叶绿体之间的运输,同时也可能参与植物对逆境胁迫的响应。     

Effects of Dryk1A,ASF and alternative splicing of CaMKⅡδ on myocardial hypertrophy in renovascular hypertensive rats

AIM: To investigate the role of dual-specificity tyrosine phosporylation-regulated kinase 1A (Dyrk1A)-alternative splicing factor (ASF)-calcium/calmodulin-dependent protein kinase Ⅱδ (CaMK Ⅱδ) pathway in the progression of myocardial hypertrophy in renovascular hypertensive rats. METHODS: The renovascular hypertension was induced by two-kidney one-clip (2K1C) method. The changes of blood pressure and myocardial hypertrophy were measured. The techniques of RT-PCR and Western blotting were used to detect CaMKⅡδ alternative splicing and the protein expression of Dyrk1A and ASF, respectively. RESULTS: Eight weeks after operation, systolic blood pressure (SBP) and diastolic blood pressure (DBP) in 2K1C rats increased (P<0.05). The increases in left ventricular weight (LVW), the ratio of LVW to body weight (BW) and the area of myocardial cells indicated that the hypertensive rats developed significant cardiac hypertrophy. The protein expression of Dyrk1A and mRNA expression of CaMKⅡδA and δB were significantly increased, while the protein expression of ASF and mRNA expression of CaMKⅡδC were decreased compared with sham-operated control rats (P<0.05). Treatment with Dryk1A inhibitor epigallocatechin gallate (EGCG) or harmine effectively attenuated cardiac hypertrophy and reversed the changes in the protein expression of Dyrk1A, ASF and alternative splicing of CaMKⅡδ (all P<0.05). CONCLUSION: Dyrk1A-ASF-CaMKⅡδ pathway plays a role in the development of myocardial hypertrophy in renovascular hypertensive rats.     

重组白介素-2增强口蹄疫病毒多表位疫苗免疫效果研究

通过研究重组白细胞介素-2（IL-2）对口蹄疫病毒（FMDV）多表位疫苗免疫效果的增强作用,为IL-2作为 FMDV 表位疫苗佐剂应用提供实验依据。6周龄雌性 Balb／c 小鼠,分为4组,第1组联合免疫表达猪 IL-2的重组腺病毒（rAd5poIL-2）和串联表达 FMDV 多表位基因的重组腺病毒（rAd5EGS）,第2、3和4组分别注射 rAd5EGS、灭活疫苗和 PBS,间隔2周免疫1次,共免疫3次。首免后每周采血分离小鼠血清,通过 ELISA 检测血清中特异性 IgG;首免后6周检测血清中抗体亚型 IgG1、IgG2a 及细胞因子 IL-4和 IFN-γ的表达水平,同时分离脾细胞,MTT 法检测淋巴细胞增殖指数。结果显示,联合免疫 rAd5poIL-2和rAd5EGS 既能增强 rAd5EGS 诱导小鼠特异性 IgG、IgG1和 IgG2a 的分泌,又能增强其诱导淋巴细胞增殖能力,且免疫效果强于灭活疫苗;细胞因子检测结果显示,联合免疫 rAd5poIL-2后能增强 rAd5EGS 诱导小鼠 IL-4和 IFN-γ的分泌,且其诱导 IL-4和 IFN-γ分泌能力也强于灭活疫苗。说明重组 IL-2能有效增强FMDV 多表位疫苗诱导抗体分泌能力和细胞免疫应答,rAd5poIL-2有望作为 FMDV 表位疫苗的候选佐剂。