73例中国人血友病甲基因突变的分析

我们用Southern blotting、PCR、变性梯度凝胶电泳(DGGE)和DNA测序等方法对73例血友病甲患者(经上海瑞金医院测定血浆FVⅢ:C和vWF:Ag诊断,其中无亲缘关系患者65例,按FVⅢ:C水平分为轻、中、重三型。FVⅢ:C< 2%为重型,共47例;FVⅢ:C 2%-5%为中型,共9例;FVⅢ:C5%-25%为轻型,共1 7例)进行FVⅢ基因突变检测。共检出内含子22倒位23例,均为重型,约占重型的49%,与国外报道相似。余下50例(其中无亲缘关系者45)用PCR-DGG E分析所有外显子及其侧翼内含子序列,发现异常条带则进行DNA测序。在17例患者中检出突变13种,其中无义突变5种,均为重型;错义突变6种,除1例外都是轻中型;小缺失2例,都是重型;其中,AA466Lys(AAG)-Thr(ACG)、719Tyr(TAC)-Stop(TAG)、AA826 Asp(GAC)-Glu(GAA)、312Ile(ATC)-xxC及AA1551-1552del(AGAA)为新发现的突变。有亲缘关系的患者都有相同的基因突变,而在无亲缘关系患者未发现相同突变。基因突变与临床表现基本相符。 Abstract:We use Southern blotting,PCR,denaturing gradient gel electrophoresis(DGGE)and DNA sequencing to detect gene mutations of haemophilia A in Chinese population.73 cases(47 severe)(FVIII:C<2%),9 moderate(FVIII:C 2%~5%),17 mild(FVIII:C 5%~25%)of haemophilia A were first screened with Southern blotting,23 were found to be the intron 22 inversion type,all being severe cases.The remaining 50 cases without intron 22 in version were examined with PCR-DGGE.Genomic DNA were amplified using GC-clamped primers covering all the exons and all flanking intron regions.Abnormal bands were sequenced.13 different mutations were identified,including 5 nonsense mutations,6 missense mutations and 2 small deletions.5 mutations,AA466Lys(AAG)-Thr(ACG),AA719Tyr(TAC)-Stop(TAG),AA826Asp(GAC)-Glu(GAA),AA312Ile(ATC)-xxC and AA1551-1552del(AGAA)have not been reported before.Generally the genetic defects correspond to the clinical conditions.     

柑桔爆皮虫卵巢发育动态

本研究利用室内饲养成虫,对柑桔爆皮虫的蛹及正常取食和交配、正常取食但未交配以及正常交配但未取食三种处理的雌成虫的卵巢发育进行了系统观察。结果显示：该虫具有1对卵巢,每侧有5根卵巢管。前两种处理的雌虫卵巢管均能正常发育,成虫寿命在30天左右; 没有取食的雌虫寿命只有7天左右,在其卵巢管的生长区和成熟区均无卵形成。根据卵巢的形状、卵的产生过程、卵巢萼内有无卵粒以及卵黄沉积情况等将卵巢发育程度分为6个级别,即发育初期（0级）、卵黄沉积前期(Ⅰ级)、卵黄沉积期(Ⅱ级)、成熟待产期(Ⅲ级)、产卵盛期(Ⅳ级)和产卵末期(Ⅴ级)。每头雌虫最高怀卵量在140粒左右,根据雌虫怀卵量变化趋势,推测正常取食和交配的柑桔爆皮虫雌虫在出孔后10天左右开始产卵,产卵历期可达22天左右。据此提出该虫卵巢管发育到成熟待产期(Ⅲ级)之前（即羽化出孔后10天之内）为出孔成虫的防治适期。     

High‐fat diet‐induced dysbiosis mediates MCP‐1/CCR2 axis‐dependent M2 macrophage polarization and promotes intestinal adenoma‐adenocarcinoma sequence

High‐fat diet (HFD) is a well‐known risk factor for gut microbiota dysbiosis and colorectal cancer (CRC). However, evidence relating HFD, gut microbiota and carcinogenesis is limited. Our study aimed to demonstrate that HFD‐induced gut dysbiosis promoted intestinal adenoma‐adenocarcinoma sequence. In clinical study, we found that HFD increased the incidence of advanced colorectal neoplasia (AN). The expression of monocyte chemoattractant protein 1 (MCP‐1), CC chemokine receptor 2 (CCR2) and CD163 in CRC patients with HFD was significantly higher than that in CRC patients with normal diet. When it comes to the Apc^min/+ mice, HFD consumption could induce gut dysbiosis and promote intestinal carcinogenesis, accompanying with activation of MCP‐1/CCR2 axis that recruited and polarized M2 tumour‐associated macrophages. Interestingly, transfer of faecal microbiota from HFD‐fed mice to another batch of Apc^min/+ mice in the absence of HFD could also enhance carcinogenesis without significant body weight gain and induced MCP‐1/CCR2 axis activation. HFD‐induced dysbiosis could also be transmitted. Meanwhile, antibiotics cocktail treatment was sufficient to inhibit HFD‐induced carcinogenesis, indicating the vital role of dysbiosis in cancer development. Conclusively, these data indicated that HFD‐induced dysbiosis accelerated intestinal adenoma‐adenocarcinoma sequence through activation of MCP‐1/CCR2 axis, which would provide new insight into better understanding of the mechanisms and prevention for HFD‐related CRC.     

两种类型聚半乳糖醛酸酶的提纯及性质

黑曲霉(AspergilluS niger)AS 3.3883所产果胶酶经DEAE Sephadex A50及Sephadex G100柱层析分离出电泳纯的两种聚半乳糖醛酸酶(PG1,PG2),并对它们的性质及结构进行了比较研究。结果证明两种酶作用的最适条件、动力学性质、分子量、氨基酸组成及金属离子对酶活力影响等方面有很大差异,但二者的每个摩尔的活力及酶的构象很相似。     

单面针的生物碱研究

自芸香科(Rutaceae)花椒属植物单面针(Zanthoxylum nitidum var. fastuosum How ex Huang)的根皮中分得五种已知生物碱:乙氧基白屈菜红碱(ethoxychelerythrine)(Ⅰ);氯化光花椒碱(nitidine chloride)(Ⅱ);去甲基白屈菜红碱(des-N-methychelerythrine)(Ⅲ);α—别隐品碱(α-allocryptopine)(Ⅳ);鹅掌揪宁(liriodenine)(Ⅴ).     

Induction of defence gene expression by oligogalacturonic acid requires increases in both cytosolic calcium and hydrogen peroxide in Arabidopsis thaliana

Responses to oligogalacturonic acid (OGA) were determined in transgenic Arabidopsis thaliana seedlings express-ing the calcium reporter protein aequorin. OGA stimulated a rapid, substantial and transient increase in the concentration of cytosolic calcium ([Ca^2 ]cyt) that peaked after ca. 15 s. This increase was dose-dependent, saturating at ca. 50 μg Gal equiv/ml of OGA. OGA also stimulated a rapid generation of H202. A small, rapid increase in H2O2 content was followed by a much larger oxidative burst, with H2O2 content peaking after ca. 60 min and declining thereafter. Induction of the oxidative burst by OGA was also dose-dependent, with a maximum response again being achieved at ca. 50 μg Gal equiv/mL. Inhibitors of calcium fluxes inhibited both increases in [Ca^2 ]cyt and [H2O2], whereas inhibitors of NADPH oxidase blocked only the oxidative burst. OGA increased strongly the expression of the defence-related genes CHS,GST, PAL and PR-1. This induction was suppressed by inhibitors of calcium flux or NADPH oxidase, indicating that increases in both cytosolic calcium and H2O2 are required for OGA-induced gene expression.     

杂交水稻及其“三系”线粒体DNA的AP—PCR指纹图谱

为了研究水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）细胞质雄性不育（ＣＭＳ）与线粒体基因组的关系,应用ＡＰ－ＰＣＲ 分析,用７ 个任意单引物对６ 种水稻品系线粒体ＤＮＡ 进行了扩增。水稻线粒体ＤＮＡ 的ＡＰ－ＰＣＲ 产物可分为三种类型：（１）所有供试品系均能扩增的片段,它们代表了线粒体ＤＮＡ 在进化上的保守性序列。有４ 个引物检测到这类片段。（２）２ 个以上水稻品系共同出现而在全部供试材料间存在差异的扩增片段,这类片段是检测水稻线粒体ＤＮＡ多态性的主要来源。（３）一种细胞质类型所特有的扩增片段,从引物Ｒ２ 和Ｖ５ 的扩增产物中发现了这类片段,它们可能与ＣＭＳ有关联。另外,ＷＡ型不育系珍汕９７Ａ 与其杂种之间在６ 个引物的扩增图谱上均存在不同程度的差异,说明两者的线粒体ＤＮＡ序列结构可能存在某种差别     

高产申嗪霉素和吩嗪-1-酰胺的水稻根际铜绿假单胞菌PA1201分离、鉴定与应用潜力

【目的】从水稻根际筛选能高效抑制水稻病原菌的细菌,分析和鉴定其形态和生化特征,为开发新型绿色农药奠定基础。【方法】从水稻根际分离能以1-氨基环丙烷-1-羧酸(ACC)为唯一碳源的菌株,根据菌株形态和生化特性、16S r DNA序列比对和磷脂脂肪酸图谱,对该菌株进行鉴定。通过氯仿萃取抽提、高效液相色谱分析,确定菌株PA1201在PPM培养基和黄豆粉培养基中申嗪霉素和吩嗪-1-酰胺的产量。【结果】菌株PA1201能有效抑制水稻纹枯病菌和水稻白叶枯病菌的生长,属于铜绿假单胞菌(Pseudomonas aeruginosa sp.PA1201);PA1201产生两种抑菌代谢产物申嗪霉素和吩嗪-1-酰胺,在PPM和黄豆粉培养基中申嗪霉素的产量可达81.7 mg/L和926.9 mg/L,吩嗪-1-酰胺的产量亦可达18.1 mg/L和489.5 mg/L;PA1201产生大量胞外蛋白酶,对人肺腺癌细胞系A549和黑腹果蝇具有一定毒性。【结论】PA1201的申嗪霉素产量比现有生产菌株的出发菌株M18高3-4倍,还能产生另一种抑菌活性更高的衍生物吩嗪-1-酰胺,具有进一步开发的潜力。     

水曲柳大孢子发生及雌配子体发育的研究

水曲柳（ＦｒａｘｉｎｕｓｍａｎｄｓｈｕｉｃａＲｕｐｒ．）子房为二心皮;二室,中轴胎座,每室内具二个倒生胚珠,薄珠心,单珠被。单个孢原细胞分化于珠心顶端表皮之下,直接发育成为大孢子母细胞,大孢子四分体为直线形,通常为合点端第一大孢子具功能,胆囊发育为蓼型。