GbLMI1 over-expression improves cotton aboveground vegetative growth

Leaves are the main organ for photosynthesis and organic synthesis in cotton.Leaf shape has important effects on photosynthetic efficiency and canopy formation,thereby affecting cotton yield.Previous studies have shown that LMI1(LATE MERISTEM IDENTITY1)is the main gene regulating leaf shape.In this study,the LMI1 gene was inserted into the 35S promoter expression vector,and cotton plants overexpressing LMI1(OE)were obtained through genetic transformation.Statistical analysis of the biological traits of the T_(1) and T_(2) populations showed that compared to the wild type(WT),OE plants had significantly larger leaves,thicker stems and significantly greater dry weight.Furthermore,plant sections of the main vein and petiole showed that the numbers of cells in those tissues of OE plants were significantly greater.In addition,RNA-seq analysis revealed the differential expression of genes related to gibberellin synthesis and NAC gene family(genes containing the NAC domain)between the OE and WT plants,suggesting that LMI1 is involved in secondary wall formation and cell proliferation,which promotes stem thickening.Moreover,Gene Ontology(GO)analysis revealed enrichment in the terms of calcium ion binding,and Kyoto Encyclopedia of Genes and Genomes(KEGG)analysis showed enrichment in the terms of fatty acid degradation,phosphatidylinositol signal transduction system,and c AMP(cyclic adenosine monophosphate)signal pathway.These results suggested that LMI1 OE plants are responsive to gibberellin hormone signals,and have altered messenger signals(c AMP,Ca^(2+))which amplify this function,to promote stronger aboveground vegetative growth.This study found the LMI1 greatly increased the vegetative growth in cotton,which is the basic requirement for higher yield.

The tumor-selective over-expression of the human Hsp 70 gene is attributed to the aberrant controls at both initiation and elongation levels of transcription

The tumor selective over-expression of the human Hsp70 gene has been well documented in human tumors,linked to the poor prognosis,being refractory to chemo-and radio-therapies as well as the advanced stage of tumorous lesions in particular.However,both the nature and details of aberrations in the control of the Hsp70 expression in tumor remain enigmatic.By comparing various upstream segments of the Hsp70 gene for each''s ability to drive the luciferase reporter genes in the context of the tumor cell lines varying in their p53 status and an immortal normal liver cell line,we demonstrated in a great detail the defects in the control mechanisms at the both initiation and elongation levels of transcription being instrumental to the tumor selective profile of its expression.Our data should not only offer new insights into our understanding of the tumor specific over-expression of the human Hsp70 gene,but also paved the way for the rational utilization of the tumor selective mechanism with the Hsp70 at the central stage fortargeting the therapeutic gene expression to human tumors.

Protocol for Artificial MicroRNA Mediated Over-Expression of miR820 in Indica Rice

In the present study, we illustrate the strategy and protocol required to generate rice transgenics over-expressing the 21-nt form of Osa-miR820. The miR exists in two size variants of 21-nt and 24-nt so the natural precursor cannot be employed for the purpose of miR over-expression as the cellular machinery can process both size variants thereby masking the role of PTGS regulation. Hence, we adopted the artificial miR technology to specifically over-express the 21-nt species in the transgenics. During the course of experiments it was observed that the amiR constructs probably interfered with the regeneration of the transformed callus, necessitating protocol modifications. The results indicate the successful over-expression of the 21-nt miR species. These plants can serve as a useful source for the functional dissection of the role played by the 21-nt Osa-miR820 species. They will also be valuable in highlighting the importance for the existence of a dual mode of miR mediated target regulation.

SIMULTANEOUS OVER-EXPRESSION OF INSULIN-LIKE GROWTH FACTOR- Ⅱ (IGF- Ⅱ ) AND IGF- Ⅱ RECEPTOR(IGF- Ⅱ R) GENES IN HUMAN PRIMARY CANCER-IMPLICATION OF AUTOCRINE AND PARACRINE MECHANISM IN AUTONOMOUS GROWTH OF HEPATIC CANCER

This is first report about the simultaneous over-expression of both Insulin-like growth factor (IGF- I ) and its receptor (IGF- I R) at mRNA level in human primary hepatic Cancer (PHC). In 10 PHC samples from China, IGF-I and IGF- I R were both over-expressed, whereas only a background signal was detected in normal liver. In 5 pairs of PHC and its non- tumorous adjacent liver tissues from South Africa, IGF- I and IGF- I R were also over-expressed in PHC. mRNA expression of IGF- I in all 5 cases and IGF- I R in 4 of 5 cases were higher in cancer than non- tumorous adjacent liver tissues. These results strongly implicate that an autocrine and/ or paracrine mechanism might be Involved in formation and progression of PHC.

外周血GLDH、GLDH/ALT水平对原发性胆汁性胆管炎患者疾病分期的诊断价值

目的探讨谷氨酸脱氢酶(GLDH)、GLDH/ALT比值对原发性胆汁性胆管炎(PBC)分期的诊断效能。方法选取已明确病理分期的PBC患者212例,检测其外周血中GLDH水平,并计算FIB-4、APRI、AAR、RPR等一系列无创性血清学指标。通过Spearman分析各指标与PBC分期的相关性,绘制受试者工作特征曲线评价其诊断效能。结果Ⅰ/Ⅱ期患者,GLDH及GLDH/ALT水平差异无统计学意义(P>0.05);Ⅱ/Ⅲ期及Ⅲ/Ⅳ期患者,GLDH及GLDH/ALT水平差异均有统计学意义(P<0.05);PBC病理分期与ALT(r=0.006,P=0.152)、GGT(r=-0.036,P=0.182)、GPRI(r=0.340,P=0.055)无明显相关性;与TBil(r=0.401,P=0.009)、DBil(r=0.403,P=0.007)、AST(r=0.217,P=0.019)、RDW(r=0.422,P<0.01)、FIB-4(r=0.774,P<0.01)、APRI(r=0.620,P<0.001)、AAR(r=0.359,P<0.01)、RPR(r=0.742,P<0.01)呈正相关;与PLT(r=-0.719,P<0.01)呈负相关;外周血GLDH(r=0.162,P=0.007)、GLDH/ALT(r=0.203,P<0.01)水平与PBC病理分期呈正相关。FIB-4对PBC各期的诊断效能最好,AUC分别为0.821、0.813、0.823;在≥S1期PBC患者中,GLDH/ALT的敏感度(93.75%)最好,≥S2期及≥S3期则为FIB-4敏感度(80.00%、93.75%)最好,而APRI和RPR则分别在≥S1/≥S2期(85.45%、92.19%)、≥S3期(88.52%)中特异度最好。多指标联合后,FIB4+APRI+AAR+GLDH+GLDH/ALT对≥S2期、≥S3期PBC的诊断效能(AUC=0.931/0.878)及敏感度(82.81%/81.67%)均有所提高,FIB4+APRI+AAR+GLDH+GLDH/ALT对≥S2期PBC诊断的特异度最高(92.73%),FIB4+GLDH+GLDH/ALT对≥S3期的特异度最高(87.5%)。结论外周血GLDH及GLDH/ALT水平对PBC疾病分期具有一定的诊断价值,并呈正相关,多种无创性血清学指标联合后诊断效能更佳。

不结球白菜L-半乳糖酸-1,4-内酯脱氢酶基因(GLDH)cDNA片段克隆与RNAi表达载体构建

根据植物中hpRNA(hairpin RNA)原理,选择GLDH基因cDNA序列同源性较低区域设计1对携带双酶切位点的特异引物,以不结球白菜‘苏州青’叶片为材料提取总RNA,利用RT-PCR技术克隆了GLDH基因片段。构建中间克隆载体,经过3次亚克隆将测序正确的GLDH基因片段分别以正向和反向两种形式插入到间隔基因YYT的两端,经鉴定已插入到植物表达载体中,成功地构建了干扰GLDH基因表达的RNAi植物双元表达载体pRNAi-GLDH,并将表达载体转化到农杆菌菌株LBA4404中,为深入研究GLDH基因功能提供有效的工具。

玉米ZmGLDH基因的克隆与表达分析

为探讨玉米L-半乳糖酸-1,4-内酯脱氢酶基因(ZmGLDH)在玉米生长发育和抗逆反应中的作用,采用RT-PCR、生物信息学、实时荧光定量PCR方法,研究了ZmGLDH基因的序列结构及其在盐胁迫下的表达特征。结果表明:ZmGLDH基因有6个外显子、5个内含子,其开放阅读框编码的多肽包含39个氨基酸残基的转运肽、42个氨基酸残基的去除肽段、505个氨基酸残基的成熟肽,在近成熟肽的N-末端有一个136个氨基酸残基组成的FAD结合结构域。盐胁迫下,ZmGLDH基因表达量上调。盐胁迫1 h,其表达量上升至对照(盐胁迫0 h)的1.20倍,盐胁迫2 h上升至对照的2.31倍,盐胁迫3 h为对照的2.20倍,盐胁迫4 h为对照的1.46倍。盐胁迫期间玉米叶片中抗坏血酸(AsA)含量的变化与ZmGLDH基因表达量的变化趋势一致,说明盐胁迫诱导的ZmGLDH基因表达水平变化是影响玉米叶内AsA含量的主要因素。

血清GLDH和AST-m在诊断几种肝脏疾病中的意义

天门冬氨酸氨基转移酶(AST)广泛分布于心脏、肝脏、骨骼肌和肾脏等组织。AST有细胞质同工酶(AST-s)和线粒体同工酶(AST-m)。谷氨酸脱氢酶(GLDH)是一种线粒体酶,主要分布于肝脏、肠、胰腺、心等器官,尤以肝脏最丰富。本文对128例不同肝病患者血清中GLDH和AST-m进行检测,研究其在不同肝脏疾病中的意义。

超表达GLDH拟南芥植株对高光胁迫的响应

研究在短时间高光胁迫下2种基因型拟南芥植株(GLDH基因超表达植株gldh236OE和哥伦比亚野生型Col)的表型、叶绿素含量、叶绿素荧光等生理数据的变化.结果表明,超表达植株的抗坏血酸表达量显著高于野生型,高光对超表达植株叶片的伤害也小于对照野生型;野生型植株叶片叶绿素含量在高光处理前后并无明显变化,而超表达植株的叶绿素含量显著降低;PSII有效光化学效率Yield和光合电子传递速率ETR呈明显降低趋势,对照野生型的降低程度更明显,说明高光胁迫使PSII结构与功能受到一定程度的损伤与破坏,而抗坏血酸含量的增加能在一定程度上缓解高光胁迫所带来的伤害.

Improving the accumulation of 18α-and 18β-glycyrrhizins by over-expressing GuHMGR, GuSQS1, and GuBAS genes in Glycyrrhiza uralensis

Objective:To study the influence of over-expression of three functional genes involved in GC biosynthetic pathway,GuHMGR,GuSQS1,and GuBAS on GC production.Methods:Three plant expression vectors were constructed and transformed into Agrobacterium tumefaciens EHA105,which were used to infect Glycyrrhiza uralensis hypocotyls explants.After induction,selection,differentiation,culture,and transplantation,12,15,and 5 regenerated plants over-expressing GuHMGR,GuSQS1,and GuBAS,were obtained,respectively.Results:RT-PCR analysis showed these transgenic regenerated G.uralensis plants had 2-6 copies of GuHMGR,GuSQS1,or GuBAS.HPLC analysis showed the contents of 18α-and 18β-GC in all transgenic regenerated samples were both higher than that in the blank control.With the increase of copy numbers of GuHMGR,GuSQS1,and GuBAS,the contents of 18α-and 18β-GC were both increased in most samples.The highest 18α-and 18β-GC contents in transgenic regenerated plants were about 3.05 times and 2.80 times higher than that in the blank control,respectively.Conclusion:Over-expression of the GuHMGR,GuSQS1,and GuBAS genes enhance the accumulation of 18α-and 18β-GC in the roots and rhizomes of G.uralensis.We hope this work can lay a foundation for the molecular breeding research of G.uralensis and improving the quality of the roots and rhizomes of G.uralensis cultivars.

血清HS-CRP GLDH CHE和Fe与肝病的相关性分析

目的:测定不同肝病患者血清超敏C反应蛋白(HS-CRP)、谷氨酸脱氢酶(GLDH)、胆碱酯酶(CHE)和Fe的活性变化,比较4个项目对不同肝疾病的诊断价值。方法:收集30例门诊健康体检者和245例不同肝疾病患者的血清标本,按临床诊断分为7组,同时测定4个项目的活性,用Spss统计软件进行统计分析。结果:不同肝疾病患者血清中HS-CRP、GLDH、CHE和Fe的活性都明显升高或降低,与对照组相比差异均有非常显著性(P<0.01);同一疾病组中,HS-CRP、GLDH、CHE和Fe的活性也不一致。结论:肝疾病患者血清GLDH、HS-CRP、CHE和Fe的活性变化,为肝细胞是否实质性损害、继发感染及肝细胞线粒体病变,提供了可靠的依据,并可对临床上治疗措施的选择和并发症的认识起指导作用。

内源抗坏血酸对水稻种子萌发及幼苗生长的影响

为了解内源抗环血酸在水稻(Oryza sativa)种子中的作用,以野生型品种‘中花11’(ZH-11)、抗坏血酸合成关键酶GLDH基因的上调(超表达)株系GO-2及下调(干涉)株系GI-2为材料,研究了抗环血酸含量对其种子萌发和幼苗生长的影响。结果表明,GO-2株系的种子萌发率比野生型ZH-11高6%,比干涉型GI-2高60%。外源添加1 mmol L–1抗坏血酸后,干涉型GI-2萌发率提高了22%,GO-2及ZH-11则没有明显增加。GO-2株系在幼苗的根长、株高、分蘖数和鲜重等指标上均高于ZH-11和GI-2。实时荧光定量测定结果表明,GO-2株系叶片的GLDH基因表达量显著上调,而GI-2株系则显著下调。这说明抗坏血酸有助于维持水稻种子活力和促进水稻种子发芽和幼苗生长。

血清谷氨酸脱氢酶检测的临床应用

目的 :观察血清谷氨酸脱氢酶在肝损害或导致肝损害相关疾病时的变化情况。方法 :以岛津CL 72 0 0全自动生化分析仪 ,采用双试剂双波长法 ,分别对 2 0 9例健康者和 187例肝、胃、胰、脑等疾病患者血清中谷氨酸脱氢酶活性进行了检测 ,作相关性检验 ,并统计有关指标。结果 :显示健康人血清中谷氨酸脱氢酶含量低 ,其参考范围为 0～ 11U/L ;肝脏疾病患者血清中谷氨酸脱氢酶的活性明显高于健康组。结论 :检测血清中谷氨酸脱氢酶的活性对肝病早期诊断、掌握病情、判断预后有着独特的临床意义 ,是诊断肝细胞病变 ,特别是坏死性肝病的重要指征。

血浆谷氨酸脱氢酶活性测定在肝胆疾病中的临床应用评价

目的：探讨血浆谷氨酸脱氢酶（glutamate dehydrogenase,GLDH）活性测定在肝脏疾病中的临床意义。方法：采用速率法检测229例肝功能异常者（急性肝损伤25例;慢性乙型肝炎60例;肝硬化47例;肝衰竭31例;胆道梗阻性疾病29例;肝癌37例）及120例健康体检者（作为对照组）的血浆GLDH活性,并与AST、ALT、ALT、GGT等常用肝胆疾病指标进行比较分析。结果：本室采用速率法测定GLDH活性的参考区间为4.4～13.3 U/L。胆道梗阻性疾病和肝癌患者血浆GLDH较对照组明显升高（P〈0.05）,胆道梗阻性疾病患者的GLDH/AST比值也明显高于对照组（P〈0.05）。血浆GLDH浓度与AST浓度呈显著的正相关（r=0.230,P〈0.001）。血浆GLDH浓度与肝损伤程度相关,随病情好转而降低。结论：胆道梗阻性疾病和肝癌患者血浆GLDH升高最为明显,GLDH可作为判断肝细胞损伤程度和疗效观察的指标。

枸杞L-半乳糖酸内酯脱氢酶基因的克隆及表达分析

以青海枸杞（Lycium barbarum L.）为模板,采用RT-PCR和RACE技术,克隆枸杞L-半乳糖酸-1,4-内酯脱氢酶（GLDH）基因序列,命名为LbGLDH。LbGLDH基因全长为2 114bp,包含一个开放读码框1 767bp（编码588个氨基酸）、5′末端序列57bp、3′末端序列290bp。LbGLDH基因核苷酸序列与番茄、马铃薯、烟草GLDH基因具有88%～90%的一致性。LbGLDH编码氨基酸序列包含GLDH蛋白酶具备的FAD-binding-4和ALO结构域。qRT-PCR分析结果显示,在枸杞不同组织中LbGLDH基因的表达、抗坏血酸含量和GLDH活性变化趋势相同,表现为在枸杞的花和果实中表达量最高,成熟叶中表达量最少。推测LbGLDH基因表达促进枸杞果实中抗环血酸含量的积累。

不同品种大豆发芽过程中抗坏血酸合成积累的比较研究

测定了鲁豆 2号、淮豆 1号、吉林 3号和楚秀等 4种不同品种大豆发芽过程中抗坏血酸含量的变化情况 ,结果表明供试大豆在发芽过程中抗坏血酸含量均显著增加 ,说明大豆发芽过程中开始了抗坏血酸的合成代谢。发芽 4天后的鲁豆 2号和淮豆 1号的抗坏血酸含量高于相同发芽条件下的吉林 3号和楚秀 ,表明不同品种发芽大豆中抗坏血酸的合成能力不同。在 4种发芽大豆中均检出半乳糖酸内酯脱氢酶 (GLDH) ,其活性受光照条件的影响 ,蓝光、白光、紫外光对GLDH活性具有促进作用 ,紫外光的促进作用最显著。在相同光强和相同时间的紫外光照射下 ,楚秀和吉林 3号的抗坏血酸增加量高于鲁豆 2号和淮豆 1号 。

混合瘤胃微生物体外利用蛋氨酸、赖氨酸的研究

为研究瘤胃微生物对瘤胃液中添加的蛋氨酸、赖氨酸的利用情况,并进一步探讨添加氨基酸对瘤胃液中尿素氮、部分酶活的影响,将采集到的来自3头装有永久性瘤胃瘘管的成年萨能山羊的瘤胃液混合后,平均分装成18个瓶,随机分成3个处理组,每组6个重复。采用6重复随机因子试验设计。第1组中每瓶添加蛋氨酸2mM(0.25mol/LMet溶液8mL),第2组中每瓶添加赖氨酸2mM(0.25mol/LLys溶液8mL),第3组中每瓶加等体积蒸馏水作为空白对照组,体外培养16h。结果表明:添加氨基酸后,混合瘤胃液中的尿素氮(UN)含量显著高于对照组(P<0.05);谷氨酸脱氢酶(GLDH)活性随着体外培养时间的延长而迅速增加,并且添加蛋氨酸组的GLDH的活性明显高于其他两组(P<0.01),而添加赖氨酸组的GLDH活性则极显著低于蛋氨酸组和对照组(P<0.01);添加的氨基酸和体外培养时间对于谷丙转氨酶(GPT)活性没有显著的影响(P>0.05);添加氨基酸组的谷草转氨酶(GOT)的活性显著低于对照组(P<0.05),但体外培养时间对各组中GOT的活性没有显著的影响。说明体外培养过程中瘤胃微生物分解赖氨酸的速率要比蛋氨酸快;添加的氨基酸使UN增加,但对GOT、GPT的活性起抑制作用;添加蛋氨酸可增加GLDH活性,但添加赖氨酸却有抑制作用。

应用ROC曲线评价血清谷氨酸脱氢酶对肝病诊断的价值

目的 :分析血清谷氨酸脱氢酶和其他几种血清酶在诊断肝脏疾病中的价值。方法 :测定了数种肝病患者及健康者GLDH的活性并与常规肝功能指标比较 ,同时进行ROC曲线分析。结果 :肝病患者GLDH活性明显高于正常人 ,急性肝炎、慢性肝炎、重症肝炎其活性随病情的加重而升高 ,病情好转时下降。血清酶AST、GGT、ALT、GLDH、ALP及LDH其ROC曲线下部面积分别为 0 983、 0 899、 0 85 1、 0 75 6、 0 732和 0 6 5 8。结论 :GLDH可作为肝病早期诊断、病情监测、病损程度及判定预后指标 ,尤其在肝损害较严重并涉及到线粒体时 。