脂肪酶和酯酶的定向进化及其应用

简单介绍了脂肪酶和酯酶的定向进化的方法和策略。重点介绍了脂肪酶和酯酶的高通量筛选方法及定向进化研究所取得的主要进展。     

Screening for High-Temperature Tolerant Cotton Cultivars by Testing In Vitro Pollen Germination, Pollen Tube Growth and Boll Retention

With radical global climate change and global warming, high temperature stress has become one of major factors exerting a major influence on crop production. In the cotton (Gossypium hirsutum L.)-growing areas of China, especially in the Yangtze River valley, unexpected periodic episodes of extreme heat stress usually occur in July and August, the peak time of cotton flowering and boll loading, resulting in lower boll set and lint yield. Breeding programs for screening high temperature-tolerant cotton germplasm and cultivars are urgent in order to stabilize yield in the current and future warmer weather conditions. In the present study, 14 cotton cultivars were quantified for in vitro pollen germination and pollen tube growth in response to temperatures ranging from 10 to 50 ℃ at 5 ℃ intervals. Different cotton genotypes varied in their in vitro pollen germination and pollen tube length responses to the different temperatures. Maximum pollen germination and pollen tube length ranged from 25.2% to 56.2% and from 414 to 682 μm, respectively.The average cardinal temperatures (Tmin, Topt, and Tmax) also varied among the 14 cultivars and were 11.8,27.3, and 42.7 ℃ for pollen germination and 11.8, 27.8, and 44.1 ℃ for maximum pollen tube length. Variations in boll retention and boll numbers per plant in field experiments were found for the 14 cotton cultivars and the boll retention and boll retained per plant on 20 August varied considerably in different years according to weather conditions. Boll retention on 20 August was highly correlated with maximum pollen germination (R2=0.84) and pollen tube length (R2=0.64). A screening method based on principle component analysis of the combination of pollen characteristics in an in vitro experiment and boll retention testing in the field environment was used in the present study and, as a result, the 14 cotton cultivars could be classified as tolerant, moderately tolerant, moderately susceptible and susceptible to high temperature.     

高效液相色谱法测定大豆乳清提取物中大豆异黄酮的含量

建立了大豆乳清提取物中大豆异黄酮含量的高效液相色谱测定方法。采用Nova-Pak C₁₈（3.9×150 mm,4 μm）色谱柱;以甲醇：0.4%磷酸=30：70（v/v）为流动相分析染料木苷和黄豆苷;流速为0.7 mL·min^-1;柱温为30℃;检测波长为260 nm。试验结果表明,大豆乳清提取物中的大豆异黄酮含量为72.5%,其组成以染料木苷和黄豆苷为主,二者比例接近1∶1,苷元型大豆异黄酮未检出。染料木苷和黄豆苷的平均回收率分别为98.1%和98.4%,相对标准偏差（RSD）分别为0.7%（n=5）和0.8%(n=5)。该方法快速、准确、重复性好。     

西洋参不同部位皂苷类成分的指纹图谱研究

以西洋参（Panax quinquefolium L.）为材料,分别取主根、须根、芦头、参皮、参芯、茎、叶、花序、果实和芽孢等不同部位为材料,以高效液相法检测其皂苷类成分。利用系统聚类分析法和中药色谱指纹图谱相似度评价系统对各部位的皂苷种类和含量进行分析,将西洋参的不同部位按照成分组成进行了分类。     

Mutational screening of the CYP26A1 gene in patients with caudal regression syndrome

BACKGROUND: The retinoic acid (RA)-catabolizing enzyme Cyp26a1 plays an important role in protecting tailbud tissues from inappropriate exposure to RA. Cyp26a1-null animals exhibit caudal agenesis and spina bifida, imperforate anus, agenesis of the caudal portions of the digestive and urogenital tracts, and malformed lumbosacral skeletal elements. This phenotype closely resembles the most severe form of caudal agenesis in humans. In view of these findings, we investigated a potential involvement of the human CYP26A1 gene in the pathogenesis of caudal regression syndrome (CRS). METHODS: Mutational screening of 49 CRS patients and 132 controls was performed using denaturing high-performance liquid chromatography and sequencing. Differences in the genotype and allele frequency of each SNP were evaluated by chi(2) analysis. The biological significance of the intronic variants was investigated by transfection assays of mutant constructs and by analysis of the splicing patterns with RT-PCR. RESULTS: Mutational screening allowed us to identify 6 SNPs, 4 of which (447 C>G, 1134 G>A, IVS 1+10 G>C, and IVS 4+8 AG>GA) are new. In addition, we describe a novel 2-site haplotype consisting of the 2 intronic SNPs. Both single-locus and haplotype analyses revealed no association with increased risk for CRS. The consequences of the 2 intronic polymorphisms on the mRNA splicing process were also investigated. Moreover, using functional and computational methods we demonstrated that both of these intronic polymorphisms affect the intron splicing efficiency. CONCLUSIONS: Our research did not provide evidence that CYP26A1 has implications for the pathogenesis of human CRS. However, the relationship between CRS risk and the CYP26A1 genotype requires further study with a larger number of genotyped subjects.     

Involvement of osteopontin upregulation on mesangial cells growth and collagen synthesis induced by intermittent high glucose

Glucose fluctuations are strong predictor of diabetic vascular complications. We explored the effects of constant and intermittent high glucose on the proliferation and collagen synthesis of cultured rat mesangial cells. Furthermore, the possible involvement of osteopontin (OPN) was assessed. In rat mesangial cells cultured in 5, 25, or 5 mmol/L alternating with 25 mmol/L glucose in the absence or presence of neutralizing antibodies to OPN, β3 integrin receptor and β5 integrin receptor, the cell proliferation, collagen synthesis, and the expression of OPN and type IV collagen were assessed. In cultured mesangial cells, treatment with constant or intermittent high glucose significantly increased [³H]thymidine incorporation in a time‐dependent manner. A modest increase was observed at 12 h, and further deteriorated afterwards, and reached the maximum incorporation at 48 h. Treatment with constant high glucose for 48 h resulted in significant increases in [³H]thymidine incorporation, cell number, [³H]proline incorporation, mRNA, and protein levels of type IV collagen and OPN compared with mesangial cells treated with the normal glucose, which were markedly enhanced in cells exposed to intermittent high glucose medium. In addition, neutralizing antibodies to either OPN or its receptor β3 integrin but not neutralizing antibodies to β5 integrin can effectively prevented proliferation and collagen synthesis of mesangial cells induced by constant or intermittent high glucose. Intermittent high glucose exacerbates mesangial cells growth and collagen synthesis by upregulation of OPN expression, indicating that glycemic variability have important pathological effects on the development of diabetic nephropathy, which is mediated by the stimulation of OPN expression and synthesis. J. Cell. Biochem. 109: 1210–1221, 2010. © 2010 Wiley‐Liss, Inc.     

高温对水稻叶片蛋白质表达的影响

为明确高温对耐热性不同水稻品种叶片蛋白质表达的影响,以耐热性不同的2个籼稻品种双桂1号(不耐热)和黄华占(耐热)为材料,分别于苗期、减数分裂期及抽穗(始穗后0—10d)和灌浆早期(始穗后11—20d)进行高温处理,之后取材并采用双向凝胶电泳技术研究高温对不同水稻品种叶片蛋白质表达的影响。结果表明,高温胁迫导致叶片中蛋白质的变化呈4种状况:新蛋白质的产生,一些蛋白质表达量上调,一些蛋白质的表达被抑制,一些蛋白质表达量下调。蛋白质表达变化在两品种以及4个处理时期的表现不同,总体表现为在热敏感品种中表达谱发生变化的蛋白质总数高于耐热品种。质谱分析表明,差异蛋白质主要涉及光合作用和信号转导,该类蛋白质在热敏感品种中表现为不表达或表达量下降,而在耐热品种则表现为有新诱导的蛋白质的产生或表达量上调,表明参与光合作用和信号转导的蛋白质在水稻耐热机制中发挥了重要作用。     

Hepatitis C Virus Experimental Model Systems and Antiviral drug Research

An estimated 130 million people worldwide are chronically infected with hepatitis C virus (HCV) making it a leading cause of liver disease worldwide. Because the currently available therapy of pegylated interferon-alpha and ribavirin is only effective in a subset of patients, the development of new HCV antivirals is a healthcare imperative. This review discusses the experimental models available for HCV antiviral drug research, recent advances in HCV antiviral drug development, as well as active research be...     

谷氨酰胺对急进高原大鼠肠道微生态影响的实验研究

目的探讨谷氨酰胺对急进高原大鼠小肠黏膜形态结构及肠道微生态的影响。方法W istar大鼠50只,随机分为5组:对照组(A组)、3848米未干预组(B组)、3848米谷氨酰胺干预组(C组)、4767米未干预组(D组)和4767米谷氨酰胺干预组(E组),每组10只,急进海拔3848米和4767米造成大鼠急性缺氧模型,检测小肠黏膜上皮细胞形态结构、肠道菌群失衡及细菌易位的变化。结果高海拔缺氧组大鼠小肠黏膜变薄、肠黏膜水肿、绒毛短缩,肠道菌群失衡显著高于对照组(P0.05),且随着海拔升高,菌群失衡更明显。不同海拔高度细菌易位率也有差异。经谷氨酰胺干预后,肠道的菌群失衡及细菌易位率与高海拔缺氧组比较差异有显著性(P0.05)。结论急进高原缺氧环境可导致小肠黏膜损伤、肠道菌群失衡及细菌易位,肠黏膜屏障破坏,且随着海拔升高而上述改变更明显。谷氨酰胺具有保护肠黏膜屏障及调节肠道菌群失衡的作用。