成酶的

Diketoreductase is a new class of enzyme, and compared to other oxidoreductase, the unique property is that it can be reduced diketone substrate to corresponding dihydroxy alcohol with high stereoselectivity.

双羰基还原酶（Diketoreductase，DKR）是一类全新的酶，与其他的氧化还原酶相比，其独特之处在于它能够同时还原两个羰基还原成羟基，并有高度的立体选择性。

Holoenzyme A catalytically active complex made up of an apoenzyme and a coenzyme.

全酶：由一个脱辅基酶蛋白和辅酶组合而成的具有催化活性的复杂化合物。

The PSCA_3 fragment was selected for its superior expression level in eukaryotic cells.Then the sig-PSCA_3-Fc-GPI genetic fragment was cloned into pVAX1-neo-IRES-GM/B7 vector to construct the final immunological inhanced DNA vaccine pVAX1-PSCA_3-FcGB. Immunofluorescence and flow cytometry were used to confirm the expression of PSCA_3 fragment by transfected into Cos7 cell.Finally,the anti-tumor effect of pVAX1-PSCA_3-FcGB was tested in murine prostate cancer model generated by RM-1 cell line.The animal was immunized with pVAX1-PSCA_3-FcGB DNA vaccine by intramuscular injection plus electroporation,pVAX1 and pVAX1-PSCA_1-FcGB plasmid were used as control.The inhibitory effect of tumor was investigated by observion of forming time,volume and inhibition ratio of tumor.Results:DNA sequencing conformed that the heterological PSCA fusion antigen fragment which was synchronized by overlapping-extending-PCR,was consistent to design.Enzyme digestion analysis showed that the 1 to 4 copies heterological PSCA fusion antigen fragments were constructed successfully.

方法(1)检索GenBank，选择包含人主要T细胞抗原表位序列的人PSCA基因片段，应用异种化抗原设计技术，保留人T细胞抗原表位，设计异种化PSCA融合抗原片段；(2)根据核酸序列按中心模板法设计引物，应用重叠延伸PCR技术拼接合成异种化PSCA融合抗原片段基因，以PCR、限制性酶切和DNA序列测定法进行鉴定：(3)利用DNA限制性内切酶BssHⅡ和MluⅠ酶切后粘端互补的特点，采用同尾酶法构建1—4拷贝异种化PSCA融合抗原片段(PNCA_1-PSCA_4)，并将上述片段分别插入真核表达载体pCI-neo-Fc-GPI中，转染293T细胞，借助免疫荧光+流式细胞术考察插入片段表达效率，最终选定PSCA_3片段进行下一步研究；(4)将sig-PSCA_3-Fc-GPI基因片段自pCI-PSCA_3-Fc-GPI质粒上切下，插入pVAX1-neo-IRES—GM/B7载体中，构建免疫增效DNA疫苗pVAX1-PSCA_3-FcGB，并应用转染Cos7细胞+免疫荧光/流式细胞术方法鉴定其在真核细胞中的表达情况；(5)给8周龄雄性C57BL/6小鼠皮下种植RM-1细胞，制备小鼠前列腺癌模型，并采用股四头肌肌肉注射+电脉冲法(Electroporation，EP)接种DNA疫苗质粒pVAX1-PSCA_3-FcGB，同时接种pVAX1空载体质粒和pVAX1-PSCA_1-FcGB质粒作为对照，通过观察计算免疫动物的成瘤时间、肿瘤体积和抑瘤率，来评价该DNA疫苗在小鼠体内的抑瘤效果。

Fat tissue is hydrolyzed and released fatty acid as well triolein by lipase; on the other hand, high fat diet can't be directly absorbed across the intestinal mucosa.it must be firstly decomposed into one 2-monoglyceride and two free fatty acids by pancreatic lipase ,which is delivered into the lumen of the gut as a constituent of pancreatic juice.

脂肪酶能够使体内的多余脂肪组织分解成三酸甘油酯，另一方面，人体摄入高脂肪食物，并不能直接被小肠粘膜吸收，而是先被小肠内的胰脂肪酶分解成2-单酸甘油酯和两个游离脂肪酸，然后和胰液一起被运输到肠道的内腔被吸收。

No.16A was treated by acetone into three groups of degumming liquor with different compositions of pectinase and-βmannanase.

No.16A发酵液中的果胶酶和甘露聚糖酶处理成不同活性比例的3组脱胶酶液。

Then, they were transferred into the mycelium culture fluid which includes 30% sucrose and 1% or 1.5% glycin under the condition of 20℃, 150 rpm for 48 h. In order to obtain protoplasts of mass and high quality, the strain F46 protoplasts given after combination use of the 10 mg·mL-1 lysozyme and 10 mg·mL-1 snailase at 34℃for 3 h; and the SC1 was treated by 10 mg·mL-1 lysozyme at 34℃for 2.5 h.

确定了双亲原生质体形成与再生的最佳条件：菌株F46、SC1采用二次菌丝培养，即在蔗糖含量20%菌丝培养液里，25℃条件下150 rpm培养48 h；然后按15%的接种量转接到蔗糖含量30%，甘氨酸含量分别为1%、1.5%的菌丝培养液中，同样条件下，继续培养48 h；在34℃条件下，菌株F46用等量混合的蜗牛酶和溶菌酶20 mg·mL-1，菌株SC1用10 mg·mL-1溶菌酶，分别酶解3 h、2.5 h，不仅有大量的原生质体形成，且再生率也高。

RESULTS AND CONCLUSION: Osteoblast was fusiformed-shaped and had plentiful processes. Nucleus was orbicular-ovate and leaning to lateral side. Soma was large, and plasma was abundant. Alkaline phosphatase staining suggested that a great number of gray-black particles were observed in plasma, and some region was darkly stained.

结果与结论：成骨细胞呈梭形，多突起；细胞核呈卵圆形，偏于一侧；胞体大，胞浆丰富；碱性磷酸酶染色可见胞浆中含有大量的灰黑色颗粒，某些部位染成黑色，定量分析细胞内的碱性磷酸酶、骨钙素水平明显高于成纤维细胞；细胞免疫组织化学染色表明其主要合成Ⅰ型胶原。

SUC2 mainly encodes sucrose invertase. The invertase could hydrolyse sucrose into glucose and fructose which are carbon source for yeast growth. When sucrose is used as the main carbon source, invertase is necessary for the growth of yeast. Accordingly, it is of great significance for yeast glycometabolism to study the sequence, transcription and regulation of sucrose invertase.

蔗糖转换酶基因(SUC2)编码蔗糖转换酶，通过降解酵母细胞外的蔗糖成果糖和葡萄糖来提供酵母生长所需的碳源，在以蔗糖为主要碳源的培养条件下，蔗糖转换酶对于酵母生长必不可少，因此研究蔗糖转换酶基因序列、转录、调控等对酵母蔗糖代谢具有重要意义。

Results of non-reducing sodium dodecyl sulphate polyacrylamide gel electrophoresis of the supernatant and aggregate precipitate formed in refolding process show that except being refolded to native egg white lysozymes, the reduced-denatured lysozymes can also form the aggregates with molecular weights being separately about 30.0 and 35.0 kD, while the reducing SDS-PAGE and the refolding results in the presence of sodium dodecyl sulphate show that these aggregates are formed chiefly through the misconnection of disulfide bonds between the reduced-denatured lysozymes, and the aggregate precipitates are formed through the non-covalent interactions between the aggregates with molecular weight being about 30.0 kD.

复性过程形成的上清和集聚体沉淀的非还原电泳结果表明，除了能复性成天然态的溶菌酶分子外，还原变性蛋白溶菌酶同时还能形成分子量分别约为30.0KD和35.0KD的蛋白溶菌酶分子集聚体；而它们的还原电泳和在SDS存在时的复性结果表明，这些集聚体主要是通过还原变性蛋白溶菌酶分子间的二硫键错配而形成，而集聚体沉淀则是通过分子量约为30.0KD的集聚体分子之间的非共价相互作用而形成。

The hFCECs were cultured by sticking tissues piece method that the cornea were divided into the limbus and central tissue piece and digestive method,respectively.For digestive method,the digestive juice of 5mg/ml DispaseⅡ+0.25%trypsin was chosed.D/F12 with 10%fetal bovine serum and 100 U/ml penicillin and streptomycin were used in this study,the culture condition was 37℃and 5%CO2,and culture medium changed every three days2、Passage of hFCECsAfter more than 80%confluenced,cells from corneal limbus hFCECs were digested for 5-15min with different concentrations of trypsin/EDTA at 37℃,and then passaged at a ratio of 1:2.The cells were subcultured on empty plates,mouse 3T3 fibroblast feeder layer,fetal corneal stromal cell feeder l ayer or HTK feeder layer respectively.And D/F12, mouse 3T3 fibroblast conditioned medium,fetal corneal stromal cells conditioned medium or HTK conditioned medium with 10%FBS and 100 U/ml penicillin and streptomycin were used respectively as the culture medium.And the culture medium were changed every three days as well.

方法一、人胎儿角膜上皮原代和传代培养1、原代培养严格无菌操作获取人胎儿角膜片，采用组织块贴壁法、酶消化法原代培养人胎儿角膜上皮细胞。2、传代培养角膜缘部的细胞生长融合达80%以上，不同稀释浓度的胰酶/EDTA消化液消化传代分别接种于空板、小鼠3T3成纤维细胞饲养层、胎儿角膜基质细胞饲养层及HTK饲养层，培养液分别采用D/F12、小鼠3T3成纤维细胞条件培养液、胎儿角膜基质细胞条件培养液、HTK条件培养液。

The split between the two groups can hardly be papered over.

这两个团体间的分歧难以掩饰。

This approach not only encourages a greater number of responses, but minimizes the likelihood of stale groupthink.

这种做法不仅鼓励了更多的反应，而且减少跟风的可能性。