In order to study the altered molecular events during laryngeal carcinogenesis and elucidate the role of Haras oncogene amplification and mutation. we have examined their profile by polymerase chain reaction (PCR) and...In order to study the altered molecular events during laryngeal carcinogenesis and elucidate the role of Haras oncogene amplification and mutation. we have examined their profile by polymerase chain reaction (PCR) and selective oligonucleotide hybridization. We analyzed the mutational status of codon 12 of Haras in 22 laryngeal carcinomas and 10 normal tissues. and found that 7 of 22 laryngeal carcinomas contained a Ha-ras mutation at codon 12. The frequency of mutation was 32%. None of the normal tissues revealed mutation. Moreover. no amplification was found in cancers when compared to the normal. Ourfindings indicated that the activated Ha-ras gene existed in laryngeal carcinoma. and activation of the Haras gene by mutation at codon 12 might play a key role in laryngeal carcinogenesis.展开更多
mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the ...mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the maternal RNAs stored in egg. These mRNAs are being degraded as the development proceeds. In some animals, such as fish and amphibian, new transcripts do not appear until the midblastula stage (midblastula transition, MBT). In other animals, for example in mouse, the zygotic genes are expressed during very early stages of development. The mRNA programmed synthesis and degradation during embryonic development controls the cell differentiation, germlayer formation and pattern formation. All these mRNA changes could be displayed side by side as cDNA band differences by mRNA differential display and the genes corresponding to these differential mRNAs could thus be obtained.展开更多
Previous studies on mammals showed that peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)played a prominent role in regulating muscle fiber type transition and composition.However,the role of P...Previous studies on mammals showed that peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)played a prominent role in regulating muscle fiber type transition and composition.However,the role of PGC-1αin chicken muscle has seldom been explored.To investigate the effect of PGC-1αon chicken skeletal muscles in this study,the PGC-1αgene was overexpressed or silenced in chicken primary myoblasts by using lentivirus,and then the effects of the PGC-1αgene overexpression and knockdown on the mRNA expression profile of genes related to myofiber type specificity were examined during fiber formation.The results showed that overexpression of PGC-1αfrom proliferation to differentiation was accompanied by the up-regulated expression of Pax7,MyoD,and CnAα,which was significantly(P<0.01)increased after one day of transfection(1 I).The enhancement of MyoG,MEF2 c,and MyHC SM expression lagged,which was improved significantly(P<0.01)after four days of transfection(1 I3 D).Overexpression of PGC-1αdecreased(P<0.01)the MyHC FWM expression after four days of transfection(1 I3 D),and it had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM during myofiber formation.The effective silence(P<0.01)of PGC-1αby lentivirus mediating short hairpin RNA(shRNA)was detected after four days of transfection(1 I3 D)in cultures,and the lack of its function in chicken primary myoblasts significantly(P<0.01)down-regulated the expression of Pax7,MyoD,CnAα,MyoG,MEF2 c,and MyHC SM,significantly(P<0.01)up-regulated the expression of MyHC FWM,and had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM.These results indicated that the role of PGC-1αin regulating the fiber type specificity of chicken skeletal muscles might be similar to that in mammals,which interplayed with key genes related to myocyte differentiation and calcineurin signaling pathway.展开更多
To investigate the molecular aspects of osteoblastic interactions with β tricalcium phosphate （β-TCP） particles, human osteoblast-like MG-63 cells were cultured with β-TCP particles at a density of 6 mg/mL cultur...To investigate the molecular aspects of osteoblastic interactions with β tricalcium phosphate （β-TCP） particles, human osteoblast-like MG-63 cells were cultured with β-TCP particles at a density of 6 mg/mL culture medium for 48 h. Then, the mRNA expression of selected genes were quantified by real-time polymerase chain reaction （PCR）, including the attachment-related genes （α integrin and actin）, the proliferation-related gene （c-jun）, and the osteoblastic markers genes （type I collagen, osteonectin, alkaline phosphatase, RUNX2 and osteoclain）. The results showed that β-TCP particles （the average size 809 nm） significantly promote the attachment and the proliferation of MG-63 cells, and slightly enhance the osteoblastic differentiation based on the analyses of the related genes expression. This study provided scientific evidences to better reveal the underlines of functions of β-TCP in bone repair.展开更多
Objective To study the effects of multi- drug resistance associated protein gene (MRP gene)overexpression on the prognosis of patients with lung cancer. methods Paraffin - embedded tissues taken fromradical resection ...Objective To study the effects of multi- drug resistance associated protein gene (MRP gene)overexpression on the prognosis of patients with lung cancer. methods Paraffin - embedded tissues taken fromradical resection of 47 cases suffering from non - small cell lung cancer (NSCLC) were determined for theexpression of MRP gene mRNA by in situ hybridization using labelled digoxigenin probes combined withimmunohistochemistry. All the patients had been followed - up from 6 months to 3 years. Results Theoverexpression of MRP gene mRNA of all the 47 lung cancer specimens was found to be obviously related withsurvival time, effects of chemotherapy, recurrence or metastases after surgery, but not related with histology, tumorsize, node metastases, TNM stages, age and sex. Conclusion MRP gene mRNA expression is correlated with theprognosis of lung cancer patients and may be regarded as an indicator to forecast the prognosis and to choose thechemotherapy for lung cancer patients.展开更多
The mRNA differential display (DDRT-PCR) technique was adopted to find out the genes related tosettlement metamorphosis development process of Ruditapes philippinarum larvae.In this study,we haveobtained three hundred...The mRNA differential display (DDRT-PCR) technique was adopted to find out the genes related tosettlement metamorphosis development process of Ruditapes philippinarum larvae.In this study,we haveobtained three hundred and forty-six amplification bands in total from pediveliger larvae,veliger larvae,eye spot larvae and post-larvae.Sixty-five out of three hundred and forty-six bands are distinctly differen-tial display from band pattern,which can be put into four groups,standing for different expression char-acters.Sixteen differential display bands were cloned,sequenced and analyzed and nine different se-quences are obtained in the study.Three sequences have higher similarity to the cDNAs deposited indatabase and three are very similar to the rDNA of other species,considered as the rDNA of Ruditapesphilippinarum.The rest three sequences are found to be novel sequences after analyzed.Their accessionnumbers are AY916799,AY916798,and AY916797 respectively.We thought the novel sequences arepossibly relevant to the early embryo development of Ruditapes philippinarum larvae and can provide somefundamental understandings that are helpful for the improvement of scallop seed raising industry.展开更多
文摘In order to study the altered molecular events during laryngeal carcinogenesis and elucidate the role of Haras oncogene amplification and mutation. we have examined their profile by polymerase chain reaction (PCR) and selective oligonucleotide hybridization. We analyzed the mutational status of codon 12 of Haras in 22 laryngeal carcinomas and 10 normal tissues. and found that 7 of 22 laryngeal carcinomas contained a Ha-ras mutation at codon 12. The frequency of mutation was 32%. None of the normal tissues revealed mutation. Moreover. no amplification was found in cancers when compared to the normal. Ourfindings indicated that the activated Ha-ras gene existed in laryngeal carcinoma. and activation of the Haras gene by mutation at codon 12 might play a key role in laryngeal carcinogenesis.
文摘mRNA differential display was established by Liang and Pardee in 1992 for the purpose of displaying the mRNA differences between two tissues. The early embryonic development in animals is primarily controlled by the maternal RNAs stored in egg. These mRNAs are being degraded as the development proceeds. In some animals, such as fish and amphibian, new transcripts do not appear until the midblastula stage (midblastula transition, MBT). In other animals, for example in mouse, the zygotic genes are expressed during very early stages of development. The mRNA programmed synthesis and degradation during embryonic development controls the cell differentiation, germlayer formation and pattern formation. All these mRNA changes could be displayed side by side as cDNA band differences by mRNA differential display and the genes corresponding to these differential mRNAs could thus be obtained.
基金supported by the National Natural Science Foundation of China(31301967)the Natural Science Foundation of Jiangsu Province,China(BK20161322)+4 种基金the projects of Key Laboratory for Poultry Genetics and Breeding of Jiangsu Province(JQLAB-ZZ-201703)the Major Breeding Programs in Jiangsu Province,China(PZCZ201728)the earmarked fund for China Agriculture Research System(CARS-41)the Independent Scientific Foundation of Public Welfare Scientific Institutes in Jiangsu Province,China(BM2018026)the Open Projects of Key Laboratory of Chicken Genetics and Breeding,Ministry of Agriculture and Rural Affairs of China(CGB-201704)。
文摘Previous studies on mammals showed that peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)played a prominent role in regulating muscle fiber type transition and composition.However,the role of PGC-1αin chicken muscle has seldom been explored.To investigate the effect of PGC-1αon chicken skeletal muscles in this study,the PGC-1αgene was overexpressed or silenced in chicken primary myoblasts by using lentivirus,and then the effects of the PGC-1αgene overexpression and knockdown on the mRNA expression profile of genes related to myofiber type specificity were examined during fiber formation.The results showed that overexpression of PGC-1αfrom proliferation to differentiation was accompanied by the up-regulated expression of Pax7,MyoD,and CnAα,which was significantly(P<0.01)increased after one day of transfection(1 I).The enhancement of MyoG,MEF2 c,and MyHC SM expression lagged,which was improved significantly(P<0.01)after four days of transfection(1 I3 D).Overexpression of PGC-1αdecreased(P<0.01)the MyHC FWM expression after four days of transfection(1 I3 D),and it had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM during myofiber formation.The effective silence(P<0.01)of PGC-1αby lentivirus mediating short hairpin RNA(shRNA)was detected after four days of transfection(1 I3 D)in cultures,and the lack of its function in chicken primary myoblasts significantly(P<0.01)down-regulated the expression of Pax7,MyoD,CnAα,MyoG,MEF2 c,and MyHC SM,significantly(P<0.01)up-regulated the expression of MyHC FWM,and had no significant impact(P>0.05)on the expression of CnB1,NFATc3,and MyHC FRM.These results indicated that the role of PGC-1αin regulating the fiber type specificity of chicken skeletal muscles might be similar to that in mammals,which interplayed with key genes related to myocyte differentiation and calcineurin signaling pathway.
文摘目的探讨大鼠孕期暴露DBP对子代睾丸mRNA差异表达的影响。方法选取清洁级SD成年鼠,雌雄比例12交配,确定怀孕后行DBP染毒,试验组灌胃剂量为500 mg/kg,同时设置对照组,每组8只;试验组老鼠染毒至哺乳期结束,饲养子代雄鼠60 d,解剖取睾丸组织,采用二代高通量测序技术对睾丸样本进行基因差异表达分析及筛选,对差异表达基因进行GO富集分析和KEGG富集分析。结果差异表达的基因共有1421个,其中,上调基因722个,下调基因699个。筛选出的差异表达基因GO功能富集分析显示,生物过程(Biological Process BP)中,差异表达基因与老化、细胞增殖、炎症反应等密切相关;细胞成分(Cellular Component CC)中,差异表达基因主要富集在内质网膜、细胞膜等部分;分子功能(Molecular Function MF)中,差异表达基因主要与钙离子结合、趋化因子活性等密切相关。KEGG富集结果显示,孕期暴露DBP对子代雄鼠的影响集中分布在TNF信号传导途径、类固醇生成、细胞黏附分子(CAMs)、醛固酮合成和分泌等通路。结论大鼠孕期DBP暴露可致雄性子代睾丸mRNA差异表达,影响生殖系统的通路主要有TNF信号传导途径、类固醇生成、细胞黏附分子(CAMs)、醛固酮合成和分泌等。
基金National Natural Science Foundation of China(No.81190133)Self-Determined and Innovation Research Funds of WUT(No.2012-IV-069)
文摘To investigate the molecular aspects of osteoblastic interactions with β tricalcium phosphate （β-TCP） particles, human osteoblast-like MG-63 cells were cultured with β-TCP particles at a density of 6 mg/mL culture medium for 48 h. Then, the mRNA expression of selected genes were quantified by real-time polymerase chain reaction （PCR）, including the attachment-related genes （α integrin and actin）, the proliferation-related gene （c-jun）, and the osteoblastic markers genes （type I collagen, osteonectin, alkaline phosphatase, RUNX2 and osteoclain）. The results showed that β-TCP particles （the average size 809 nm） significantly promote the attachment and the proliferation of MG-63 cells, and slightly enhance the osteoblastic differentiation based on the analyses of the related genes expression. This study provided scientific evidences to better reveal the underlines of functions of β-TCP in bone repair.
文摘Objective To study the effects of multi- drug resistance associated protein gene (MRP gene)overexpression on the prognosis of patients with lung cancer. methods Paraffin - embedded tissues taken fromradical resection of 47 cases suffering from non - small cell lung cancer (NSCLC) were determined for theexpression of MRP gene mRNA by in situ hybridization using labelled digoxigenin probes combined withimmunohistochemistry. All the patients had been followed - up from 6 months to 3 years. Results Theoverexpression of MRP gene mRNA of all the 47 lung cancer specimens was found to be obviously related withsurvival time, effects of chemotherapy, recurrence or metastases after surgery, but not related with histology, tumorsize, node metastases, TNM stages, age and sex. Conclusion MRP gene mRNA expression is correlated with theprognosis of lung cancer patients and may be regarded as an indicator to forecast the prognosis and to choose thechemotherapy for lung cancer patients.
基金the National High Technology Research and Development Programme of China(No.2002AA603015)
文摘The mRNA differential display (DDRT-PCR) technique was adopted to find out the genes related tosettlement metamorphosis development process of Ruditapes philippinarum larvae.In this study,we haveobtained three hundred and forty-six amplification bands in total from pediveliger larvae,veliger larvae,eye spot larvae and post-larvae.Sixty-five out of three hundred and forty-six bands are distinctly differen-tial display from band pattern,which can be put into four groups,standing for different expression char-acters.Sixteen differential display bands were cloned,sequenced and analyzed and nine different se-quences are obtained in the study.Three sequences have higher similarity to the cDNAs deposited indatabase and three are very similar to the rDNA of other species,considered as the rDNA of Ruditapesphilippinarum.The rest three sequences are found to be novel sequences after analyzed.Their accessionnumbers are AY916799,AY916798,and AY916797 respectively.We thought the novel sequences arepossibly relevant to the early embryo development of Ruditapes philippinarum larvae and can provide somefundamental understandings that are helpful for the improvement of scallop seed raising industry.
