Association between ERK5 -322G/T polymorphism and genetic susceptibility to sporadic colorectal cancer

AIM: To investigate the correlation between extracellular signal-regulated kinase 5 ( ERK5 ) -322G/T polymorphism (rs3866958) and the susceptibility to sporadic colorectal cancer (CRC) in southern Chinese population. METHODS: ERK5 -322G/T genotypes were determined by Taqman-MGB probes in 835 CRC cases and 908 healthy controls. RESULTS: No significance of ERK5 -322G/T genotype distribution between CRC patients and controls was observed, but -322G/T decreased the susceptibility to CRC in fat people whose BMI was ≥ 24 kg/m². Compared to GG genotypes, the carriers with GT and TT genotypes had a significant decrease in the risk of CRC(OR=0.576,95%CI 0.413-0.804, P<0.01). CONCLUSION: ERK5 -322G/T polymorphism (rs3866958) has no significant relevance with sporadic CRC susceptibility, but decrease, the risk of CRC in people with fatness. The T variant genotype is an independent protective factor against sporadic CRC of overweight patients in southern Chinese population.     

IQGAP1 is overexpressed and interacts with ERK in human myeloma cells

AIM:To explore the role of IQ motif-containing GTPase-activating protein 1 (IQGAP1) in the cell proliferation of multiple myeloma and its mechanism. METHODS:RT-PCR and Western blotting were performed to detect the expression of IQGAP1 in human myeloma cell lines RPMI8226 and U266. shRNA-expressing plasmids were used to transfect into RPMI8226 cells to knock down the expression of IQGAP1. MTT assay was used to examine the proliferation of RPMI8226-shIQGAP1 (clone 1) cells, RPMI8226-shRNA negative control cells and un-transfected RPMI8226 cells with or without VEGF/IL-6 treatment. The protein levels of p-ERK1/2, ERK1/2, AKT, p-AKT, STAT3 and p-STAT3 in RPMI8226-shIQGAP1 (clone 1) cells, RPMI8226-shRNA negative control cells and un-transfected RPMI8226 cells were measured by Western blotting. The interaction between IQGAP1 and ERK was determined by the method of co-immunoprecipitation. RESULTS:IQGAP1 was overexpressed in human myeloma cell lines RPMI8226 and U266 as compared with normal lymphocytes. Transfection with shRNA targeting to IQGAP1 decreased the expression levels of IQGAP1 in RPMI8226 cells. The proliferation of RPMI8226 cells decreased when IQGAP1 was knocked down by shRNA with or without VEGF/IL-6 treatment. IQGAP1 affected the proliferation of RPMI8226 cells by regulation of MAP kinase (ERK1/2) pathway. The level of p-ERK1/2 in RPMI8226-shIQGAP1 (clone 1) cells decreased by 70.2% as compared with RPMI8226-shRNA negative control cells and un-transfected RPMI8226 cells. The interaction between IQGAP1 and ERK in RPMI8226 cells was observed. CONCLUSION: IQGAP1 plays an important role in the cell proliferation of multiple myeloma via MAP kinase (ERK) pathway.     

日粮蛋白质水平对梅花鹿瘤胃内pH值、NH_3-N和VFA浓度的影响

选用4头装置永久性瘤胃瘘管的成年梅花鹿,用4种含有不同蛋白质水平的日粮,按4×4拉丁方试验设计,对瘤胃内pH值、氨态氮(NH_3—N)和总挥发性脂肪酸(TVFA)浓度进行了测定,并研究了它们的动态变化规律以及VFA组分百分率。试验结果表明,在饲根精粗比为30:70条件下,日粮蛋白质水平对瘤胃内pH值影响较小(P>0.05);对NH_3—N浓度影响较大(P<0.05),随着日粮蛋白质水平的提高,瘤胃内NH_3—N浓度也随之升高;对TVFA浓度也有一定的影响,但各日粮间差异不显著(P>0.05)。通过VFA的组分分析发现,各日粮组间C_2/C_3比值基本接近,即它们的发酵类型基本相同。     

Evaluation of the Lime-Cooking and Tortilla Making Properties of Quality Protein Maize Hybrids Grown in Mexico

Eleven experimental and three commercial white quality protein maize (QPM) hybrids and two regular endosperm controls were planted at Celaya, Guanajuato, Mexico with the aim of comparing grain physical characteristics, protein quality, lime-cooking and tortilla making properties. All genotypes were planted under irrigation using a density of 80,000 plants/ha and fertilized with 250 kg N-60 P-60 K per hectare. When compared with the controls these QPM genotypes had lower test (77.4 vs. 76.5 kg/hL) and 1,000 kernel weights (327 vs. 307 g), softer endosperm texture (2.5 vs. 1.8 where 1 = soft, 2 intermediate and 3 hard endosperm), lower protein (10.0 vs. 8.0%), higher nixtamal water uptake after 30 min lime-cooking (50.0 vs. 53.1% moisture) and lower pericarp removal scores. The lower thousand-kernel weight and softer endosperm texture observed in the QPM genotypes lowered the optimum lime-cooking time as estimated with regression equations. Most QPM genotypes had higher amounts of lysine, tryptophan and albumins/globulins when compared with the controls. QPMs HEC 424973, HEC 774986 and HEC 734286 had the best grain traits for nixtamalization and therefore the best potential for industrial utilization. The commercial use of these QPM hybrids should benefit Mexicans who depend on tortillas as the main staple.     

Distribution of protein disulfide isomerase during maturation of pig oocytes

Oocyte maturation in mammals is characterized by a dramatic reorganization of the endoplasmic reticulum (ER). In mice, the ER forms accumulations in the germinal vesicle (GV) stage and distinctive cortical clusters in metaphase II (MII) of the oocyte. Multiple evidence suggests that this ER distribution is important in preparing the oocyte for Ca²⁺ oscillations, which trigger oocyte activation at fertilization. In this study, we investigated the time course and illustrated the possible functional role of ER distribution during maturation of porcine oocytes by immunostaining with protein disulfide isomerase (PDI). PDI forms clusters in the cytoplasm of oocytes. After immunostaining, PDI clusters were identified throughout the cytoplasm from the GV to metaphase I (MI) stage; however, at the MII stage, the PDI formed large clusters (1–2 µm) in the animal pole around the first polar body. PDI distribution was prevented by bacitracin, a PDI inhibitor. Our experiments indicated that, during porcine oocyte maturation, PDI undergoes a dramatic reorganization. This characteristic distribution is different from that in the mouse oocyte. Moreover, our study suggested that formation of PDI clusters in the animal pole is a specific characteristic of matured porcine oocytes.     

家蚕核型多角体病毒ODV囊膜蛋白基因的融合表达及引导外源蛋白进入多角体的研究

为了探索家蚕核型多角体病毒(BmNPV)的包涵体衍生病毒(ODV)囊膜蛋白在病毒粒子包涵入多角体过程中的作用,选择2个ODV囊膜蛋白ODV-E25和ODV-E56作为研究对象,利用能够表达多角体蛋白的BmNPV Polh+Bac-to-Bac表达系统构建了融合表达E25-EGFP和E56-EGFP的重组病毒,在家蚕卵巢培养细胞系(BmN)中进行表达和定位分析。结果表明2种融合蛋白均在BmN细胞中成功表达,并于荧光显微镜下观察到绿色荧光主要分布在细胞核中。从感染的BmN细胞中收集纯化多角体,观察到多角体也能激发出绿色荧光,用Western blot方法进一步证实多角体中含有融合蛋白。这一现象表明当囊膜蛋白基因与外源基因融合表达时,外源融合蛋白能够进入多角体内部,推测这2种ODV囊膜蛋白不仅在病毒粒子包涵入多角体的过程中起信号引导作用,并能引导外源目的蛋白进入多角体。     

Characterization of mutations in the two-component histidine kinase gene that confer fludioxonil resistance and osmotic sensitivity in the os-1 mutants of Neurospora crassa

Osmotic-sensitive (os-1) mutant alleles in Neurospora crassa exhibit resistance to dicarboximides, aromatic hydrocarbons and phenylpyrroles. We have previously reported that the os-1 mutants can be classified into two groups based on their resistance to fungicides and osmotic stress: type I, which are highly resistant to iprodione and fludioxonil but moderately sensitive to osmotic stress, and type II, which are highly sensitive to osmotic stress but moderately resistant to fungicides. To explain the mechanism of resistance to these fungicides, we cloned and sequenced the mutant os-1 genes that encode putative osmo-sensing histidine kinase. Within the os-1 gene product (Os1p), the type I strains, NM233t and Y256M209, carried a stop codon at amino acid position 308 and a frameshift at amino acid position 294, respectively. These mutation sites were located on the upstream of histidine kinase and the response regulator domains of Os1p, strongly suggesting that type I strains are null mutants. The null mutants, NM233t and Y256M209, were highly resistant to iprodione and fludioxonil; thus Os1p is essential for these fungicides to express their antifungal activity. The amino acid changes in Os1p, 625Pro from Leu, 578Val from Ala, and 580Arg from Gly were found in the type II strains, M16, M155-1 and P5990, respectively. Os1p is novel in having six tandem repeats of 90 amino acids in the N terminal. Each amino acid change of the type II strains was located on the fifth unit of six tandem repeats. Type II strains with single amino acid changes were more sensitive to osmotic stress than the null mutants (type I), indicating that the amino acid repeats of Os1p were responsible for an important function in osmo-regulation.     

荔枝霜疫霉中双组分信号转导系统的鉴定与表达分析

 荔枝霜疫霉(Peronophythora litchii)隶属于茸鞭生物界卵菌门疫霉属,由其引起的荔枝霜疫病是目前荔枝生产上一种最重要的病害,严重影响荔枝产量和鲜果品质。双组分信号途径在微生物中参与多种生命活动,但是在卵菌中尚未有相关研究。本研究通过生物信息学分析在荔枝霜疫霉中鉴定到2个杂合型组氨酸激酶(PlHK1、PlHK2)和1个响应调控蛋白(PlRR1)。其在卵菌中是保守存在的,并且与真菌在进化上相对独立。功能域分析表明,PlHK1和PlHK2的C端额外的融合了1个磷酸转移功能域(Hpt),这与真菌和植物的存在显著差异。转录分析表明3个基因在荔枝霜疫霉侵染阶段上调表达,并且响应渗透胁迫和氧化胁迫。以上结果揭示了双组分信号系统可能在疫霉致病过程中发挥重要作用。     

近红外反射光谱(NIRS)技术分析奶粉品质的研究

奶粉中蛋白质和脂肪是影响奶粉营养品质的主要因素,利用近红外反射光谱分析技术对来自国内不同地区的奶粉共900份样品进行蛋白质和脂肪成分测定分析。研究了不同的样品数日、光谱预处理和散射校正技术对发展奶粉近红外测定定标模型的影响。结果表明．在样品数目为200—400范围内建立的定标分析模型较理想;数学预处理中以一阶导数较好,且以“1,4,4,1”的处理组合最为理想;光谱散射校正中采用“标准正态变量转换(SNV) 趋势变换法(De—trending)”的组合建立回归方程效果较好。利用改进最小二乘法回归技术(Modified PLS)建立多种定标模型,并进行交叉验证(cross—Validation)来分析各种因素对定标模型的影响。同时筛选出较理想的蛋白质和脂肪定标分析回归方程,其中蛋白质和脂肪含量的相关系数高速0．973和0．850。探讨了NIRS技术在建模应用中的一些影响因素,以及由NIRS技术建立奶粉分析模型用于快速分析和在线检测的可行性。     

Influence of dietary lysine level on whole-body protein turnover, plasma IGF-I, GH and insulin concentration in growing pigs

Four growing pigs (initial liveweight 25.9 ± 0.54 kg, final liveweight 43.0 ± 1.06 kg) were used to study the effect of dietary lysine level on nutrient digestibility, whole-body protein turnover, plasma insulin-like growth factor-I (IGF-I), growth hormone (GH), insulin, glucose, and urea nitrogen (PUN). Four diets, containing 7.0 g (L1), 9.5 g (L2), 12.0 g (L3) and 14.5 g (L4) lysine per kg diet respectively, were formulated as experimental treatments. The animals and diets were allocated in a 4 × 4 Latin square design. Nitrogen (N) metabolism and whole-body protein turnover were measured by classical method and single-dose ¹⁵N end-product method, respectively. The blood samples were taken at the end of each experimental period. Results showed that N retention (NR) and N biological value (NBV) were significantly increased from L1 to L4 (P < 0.05). However, differences in NR and NBV between L2, L3 and L4 were not significant (P > 0.05). There was no significant difference on dry matter (DM) digestibility, organic matter (OM) digestibility and N digestibility between different treatments (P > 0.05). Whole-body protein synthesis, protein degradation and protein accretion increased markedly from L1 to L2 (P < 0.05), but did not increase further from L2 to L4. Whole-body protein accretion (y, g/kg W^0.75/d) increased with dietary lysine (x, g/kg) in a quadratic manner: y = − 0.09x² + 2.12x − 5.14 (r² = 0.96, n = 4, P < 0.05).The results also showed that differences in plasma IGF-I, GH, glucose and PUN concentration between different treatments were not significant (P > 0.05). Plasma insulin concentration (y, μIU/ml) was increased with dietary lysine (x, g/kg) in a quadratic manner: y = 0.23x² − 4.10x + 32.25 (r² = 0.99, n = 4, P < 0.05), but it was not found that plasma insulin concentration was related to NR. A significant correlation was found between NR (y, g/d) and plasma IGF-I (x, ng/ml): y = − 3.1 × 10^− 3x² + 1.31x − 122.28 (r² = 0.99, n = 4, P < 0.05).It was concluded that dietary lysine level had a significant influence on NR and whole-body protein turnover but not on plasma IGF-I and GH concentration. Plasma IGF-I may be an important factor controlling N metabolism of growing pigs. Further research was needed to study the mechanism.