miR-122靶向调控鸡BZW2基因的表达

miR-122(MicroRNA-122)是在肝脏中高表达的miRNA,在肝脏的生长发育和脂类代谢等过程发挥重要作用,BZW2 (basic leucine zipper and W2 domains 2)主要参与蛋白质合成代谢,本研究目的是确定BZW2是否为miR-122调控的靶基因.通过生物信息学预测miR-122的靶基因,并分析BZW2的3-UTR区域与miR-122种子区的配对情况,再用双荧光素酶报告系统和突变实验证明miR-122作用于BZW2的3'UTR.用荧光定量PCR检测转染miRNA-122mimic的鸡(Gallus gallus)肝癌细胞系(leghorn hepatocellar,LMH)细胞和转染LNA-antimiR-122的鸡原代肝细胞中BZW2的表达.鸡BZW2的3'-UTR区域与miR-122种子区互补配对,荧光素酶报告基因和突变实验分析表明,miR-122能够通过与BZW23'-UTR区结合抑制基因的表达;将miR-122在LMH细胞中过表达后,发现BZW2 mRNA表达水平显著下降;利用LNA-antimiR-122抑制鸡肝脏原代细胞中的miR-122后,BZW2 mRNA表达水平呈显著性上升.结果证明BZW2是miR-122的靶基因,其在mRNA水平上受到miR-122的负性调控,本研究为揭示miR-122在鸡肝脏中的广泛的功能提供了理论依据.     

两系法超级杂交稻粤杂122动态株型结构研究

对两系法超级杂交稻粤杂122苗期、分蘖盛期、第二次枝梗原基分化期、抽穗期、成熟期的株型结构进行调查分析表明,粤杂122与对照培杂67比较,生育前、中期具有明显的生长优势,表现秧苗素质好,植株生长迅速,出叶速度快,分蘖优势强,功能叶叶片较长且窄,叶片开张角较大、叶面积空间较大;生育后期则表现为最后3片功能叶叶片较长且窄、剑叶直立,有效穗数较多,穗数、粒数、粒重较均匀,千粒重较大等特点。认为上述形态性状可作为两系法超级杂交稻的株型结构特征,在育种和栽培实践中加以应用;粤杂122不同生育时期株型结构的性状值可作为华南两系超级杂交稻动态株型结构育种与栽培实践中,不同生育期形态性状选汰以及群体结构调控的参考指标。     

高产优质抗病杂交籼稻新组合天优122的选育与应用

天优122是广东省农科院水稻所以自选野败型优质籼稻不育系天丰A与自选的抗稻瘟病恢复系广恢122配组育成的杂交稻新组合,2005年2月通过广东省农作物品种审定。该组合具有高产稳产、稻米品质优、稻瘟病抗性强和生态适应性广等特性,适合华南早晚季种植和南方稻区作晚稻种植。     

酯酶同工酶电泳鉴定天优122、培杂双七种子纯度研究

利用聚丙烯酰胺凝胶电泳法对水稻秧苗针叶期的酶粗提液进行酯酶同工酶(EST)电泳分析结果表明:天优122不但具备父母本互补酶带EST3、Est4,同时也具备父本酶带Est7及母本酶带Est11、Est12,F1与双亲的酯酶酶带强弱有差异;而培杂双七不但具备父母本互补酶带Est3、Est4,同时也具备父本酶带Est8、Est9、Est10、Est11和Est12,F1与双亲的酯酶酶带强弱也有差异,说明利用酯酶同工酶鉴定杂交稻品种天优122、培杂双七的种子纯度是可行的。     

Time-course changes in the expression levels of miR-122, -155, and -21 as markers of liver cell damage,inflammation, and regeneration in acetaminophen-induced liver injury in rats

Drug-induced liver injury (DILI) is a significant threat to patient health and a major concern during drug development. Recently, multiple circulating microRNAs (miRNAs) have been reported to be potential biomarkers for DILI. To adapt and validate miRNAs for clinical use, we investigated the time-course changes in miR-122 expression levels in an acetaminophen-induced liver injury model in rats. In addition, miR-155 and miR-21 were evaluated as makers of inflammation and regeneration, respectively, to characterize liver status. Our results revealed that miR-122 is an early and sensitive biomarker of hepatocellular injury at a stage when alanine transaminase, aspartate transaminase, and total bilirubin were not detectable. However, no significant differences in the expression levels of other miRNAs (miR-155 and -21) were observed between treatment and vehicle groups. Collectively, these time-course changes in the expression levels of miRNAs may be useful as markers for clinical decision-making, in the diagnosis and treatment of DILI.     

高产、抗病、优质、早熟抗虫棉新品种冀122的选育

利用常规早熟棉品种省早441和转基因抗虫棉GK 12杂交世代选择育成的“冀122“,突出表现抗棉铃虫、高产、抗病和早熟,2003～2004年河北省冀东早熟棉组区域试验霜前皮棉较对照品种新棉33B和DP99B分别增产23.1%和17.7%,属抗枯萎病、抗黄萎病品种,适宜冀东早熟棉区及同类生态区种植.     

弱感光型高产杂交水稻新组合金稻优122的选育及应用

金稻优122是广东省农业科学院水稻研究所用自育不育系金稻13A与自选的抗稻瘟病恢复系广恢122配组育成的弱感光型三系杂交稻新组合,2008年1月通过广东省品种审定.该组合高产稳产,2005和2006年晚造参加广东省区试,分别比对照博优122增产8.59%和5.95%,均达显著水平,适宜广东、广西作晚稻种植.     

Detection of miR-122 expression in serum by Taqman probe real-time fluorescence quantitative PCR and its clinical significance

AIM: To detect the serum level of miR-122 expression by the technique of Taqman probe real-time fluorescence quantitative PCR for identifying its clinical significance. METHODS: The stem-loop RT primer, PCR primer and Taqman probe of miR-122 and U6 snRNA were designed. The expression of miR-122 in the serum samples of 27 cases of preoperative hepatocellular carcinoma (HCC), 15 cases of hepatitis B, 15 cases of hepatitis C, 15 cases of healthy control (HC), 11 cases of postoperative hepatocellular carcinoma (PHCC) and 10 cases of recurrence after postoperative hepatocellular carcinoma was detected by Taqman probe real-time fluorescence quantitative PCR. U6 snRNA was used as the internal control. RESULTS: The results showed that the method of Taqman probe real-time fluorescence quantitative PCR could detect the amplification signal of serum miR-122. The expression level of serum miR-122 in the patients with HCC, hepatitis B, hepatitis C and recurrence was higher than that in HC and the patients with PHCC. Meanwhile, the serum level of miR-122 in the patients with hepatitis C was higher than that in the patients with HCC, hepatitis B and recurrence. However, no difference of miR-122 expression level among HCC, hepatitis B and recurrent patients was observed. The miR-122 level was lower in PHCC patients than that in HCC and recurrent patients. In hepatitis B virus surface antigen (HBsAg) and/or hepatitis B virus e antigen (HBeAg) positive patients, the miR-122 level was higher than that in the negative ones. The miR-122 level in hepatitis C antibody (HCV-Ab) positive patients was raised compared with the negative ones. The serum level of alanine aminotransferase (ALT) was positively correlated with the serum level of miR-122 (r=0.34, P<0.05). The miR-122 expression level in the patients with serum AFP≥400 μg/L was higher than that in the patients with serum AFP<400 μg/L. CONCLUSION: The method of Taqman probe real-time fluorescence quantitative PCR can detect the serum level of miR-122 expression. Serum miR-122 might be used as a new biomarker of liver diseases, especially in the early diagnosis of primary hepatocellular carcinoma, the curative effect of surgical operation and the prognosis.     

miRNA-122 promotes mouse embryonic stem cell-derived hepatic precursor cells to differentiate into hepatocytes

AIM：To investigate whether miRNA-122 (miR-122) promotes the differentiation of mouse embryonic stem cell (ESC)-derived hepatic precursor cells (HPCs) into hepatocytes. METHODS：Mouse ESCs were initially induced to differentiate into HPCs by stimulating with fibroblast growth factor 4 (FGF-4), sodium butyrate and dexamethasone (Dex) sequentially. Then a recombinant adenovirus expressing vector pAV.Ex1d-CMV>miR-122/IRES/eGFP was constructed by Gateway technology and transfected into the mouse ESC-derived HPCs 9 d after induction, so as to gain the cells with stable high expression of miR-122. The morphological changes of transfected cells were observed under inverted phase-contrast microscope. The liver-specific gene expression levels were detected by real-time RT-PCR. The liver-specific protein expression levels were also detected by immunofluorescence method. The liver functions were assessed by indocyanine green (ICG) uptake experiment, glycogen staining and urea synthesis function test. RESULTS：The mouse ESCs were successfully induced into HPCs by stimulating with FGF-4, sodium butyrate and Dex sequentially. At 6 d after transfection of miR-122, the morphology of the cells was closer to the mature hepatocytes. The mRNA levels of liver-specific genes such as albumin (ALB), transthyretion, α1-antitrypsin, glucose-6-phosphatase, cytokeration 8, cholesterol 7α-hydroxylase and cytochrome P450 3A4 were up-regulated. The expression levels of liver-specific proteins such as ALB and cytokeratin 18 were increased, while alpha-fetoprotein was decreased. The results of ICG uptake experiment, glycogen staining and urea synthesis function test indicated that the hepatocyte functions were strengthened as compared with control group. CONCLUSION：The combination of FGF-4, sodium butyrate and Dex successfully induces the mouse ESCs into HPCs. Over-expression of miR-122 effectively promotes the differentiation and maturation of mouse HPCs.     

优质高产杂交粳稻新组合新8优122的选育与应用

用新8A与爪哇型恢复系GR03122配组育成的三系杂交中粳新组合新8优122,具有优质、高产稳产、耐肥抗倒、抗病性好等优点,2008年通过安徽省农作物品种审定委员会审定。