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AP2M1抑制弥漫性大B细胞淋巴瘤细胞增殖和侵袭
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作者 李瑗春 严学倩 +3 位作者 范丹 及月茹 肖方 高静 《基础医学与临床》 2024年第3期308-316,共9页
目的探究接头相关蛋白质复合体2亚基μ1(AP2M1)对弥漫性大B细胞淋巴瘤(DLBCL)细胞增殖和侵袭的调控作用。方法将人弥漫性大B细胞淋巴瘤细胞系OCI-LY8分为对照组、NC-LV组、AP2M1-LV组。用Lipofectamine 2000进行细胞转染。四甲基偶氮唑... 目的探究接头相关蛋白质复合体2亚基μ1(AP2M1)对弥漫性大B细胞淋巴瘤(DLBCL)细胞增殖和侵袭的调控作用。方法将人弥漫性大B细胞淋巴瘤细胞系OCI-LY8分为对照组、NC-LV组、AP2M1-LV组。用Lipofectamine 2000进行细胞转染。四甲基偶氮唑盐(MTT)法检测细胞增殖,流式细胞仪检测细胞凋亡,Transwell小室法检测细胞迁移和侵袭。Western blot检测AP2M1、表皮生长因子受体(EGFR)、p-磷脂酰肌醇3激酶(PI3K)和p-蛋白质激酶B(AKT)蛋白表达。结果与对照组相比,AP2M1-shRNA组细胞中AP2M1的mRNA和蛋白相对表达量均降低(P<0.05),相对细胞活力升高(P<0.05),细胞凋亡率降低(P<0.05),迁移和侵袭细胞数量均升高(P<0.05);EGFR的蛋白相对表达量及PI3K和AKT的磷酸化水平均升高(P<0.05)。与对照组相比,AP2M1-LV组细胞中AP2M1的mRNA和蛋白相对表达量均升高(P<0.05),相对细胞活力降低(P<0.05),细胞凋亡率升高(P<0.05),迁移和侵袭细胞数量均降低(P<0.05),EGFR的蛋白相对表达量及PI3K和AKT的磷酸化水平均降低(P<0.05)。结论AP2M1的过表达部分通过抑制EGFR/PI3K/AKT信号通路来抑制DLBCL细胞的增殖和侵袭。 展开更多
关键词 弥漫性大B细胞淋巴瘤 接头相关蛋白质复合体2亚基μ1(ap2M1) 表皮生长因子受体 磷脂酰肌醇3激酶 蛋白质激酶B
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Comparison of 2-Acetyl-1-Pyrroline (2AP) in Rice Leaf at Different Growth Stages Using Gas Chromatography
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作者 Manoch Kongchum Dustin L. Harrell Steve D. Linscombe 《Agricultural Sciences》 2022年第2期165-176,共12页
Aromatic rice lines were examined for 2-Acetyl-1-Pyrroline (2AP) content in leaf tissue at five different growth stages (tillering, panicle initiation, 50% heading, booting, and maturity). A small plot trial with plot... Aromatic rice lines were examined for 2-Acetyl-1-Pyrroline (2AP) content in leaf tissue at five different growth stages (tillering, panicle initiation, 50% heading, booting, and maturity). A small plot trial with plot size of 1.42 m × 4.88 m (7 row-plots) was arranged in completely randomize design with three replications. Dry-seeded, delayed flood cultural practice was used in this study. The experiment was conducted at three locations. The average 2AP concentrations in leaf tissue at tillering stages were higher than the other four growth stages. 2AP levels were declined when rice plant reached booting. AP levels decreased slightly at heading stage and decreased significantly at maturity. There was no significant different between 2AP in leaf at 50% heading from three locations as well as the 2AP content in rice grain. Correlations between 2AP in leaf and 2AP in grain were significantly in all five growth stages. The highest correlation coefficient was found between 2AP in leaf at booting and grain (r = 0.811**) and lowest was in the leaf at harvest (r = 0.564**). Results indicated that 2AP could be determined in leaf tissue at early growth stage. 展开更多
关键词 2-acetyl-1-pyrroline (2ap) Aromatic Rice Leaf Tissue Gas Chromatography
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AP2S1在膀胱癌组织中的表达及其临床意义
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作者 王明 张岩 王超奇 《现代肿瘤医学》 CAS 北大核心 2023年第12期2284-2288,共5页
目的:探索衔接因子相关蛋白复合体2σ1(adaptor-related protein complex 2,sigma 1 subunit,AP2S1)在膀胱癌组织中的表达水平及相关临床意义。方法:基于cBioportal数据库平台下载和整理了膀胱尿路上皮癌(bladder urothelial carcinoma,... 目的:探索衔接因子相关蛋白复合体2σ1(adaptor-related protein complex 2,sigma 1 subunit,AP2S1)在膀胱癌组织中的表达水平及相关临床意义。方法:基于cBioportal数据库平台下载和整理了膀胱尿路上皮癌(bladder urothelial carcinoma,BLCA)数据集相关数据,同时基于TCGA患者的基因表达谱和临床病理特征数据,我们分析了AP2S1与BLCA患者临床病理特征及生存预后的相关性。结果:免疫组化分析显示AP2S1在BLCA组织中的表达明显高于正常组织,有统计学意义(P<0.05)。AP2S1在BLCA组织中的阳性表达率为72.9%(70/96),对照组癌旁正常组织中AP2S1阳性表达率为25.0%(7/28)。临床样本分析结果显示:膀胱肿瘤大小、临床分期不同、病理级别高低、有无疾病复发,与AP2S1的表达相关且有统计学意义;患者性别、年龄及有无淋巴血管浸润与AP2S1表达水平无相关性。<3 cm亚组分析发现AP2S1阳性表达率为60.00%(24/40),≥3 cm亚组为82.14%(46/56);≤T_(1)期亚组分析发现AP2S1阳性表达率为67.14%(47/70),≥T_(2)期组为88.46%(23/26);在病理分级为高级别组中其阳性表达率为81.82%(45/55),低级别组为60.98%(25/41);无淋巴血管浸润组其阳性表达率为70.13%(54/77),有淋巴血管浸润组为84.21%(16/19);在无疾病复发组其阳性表达率为60.00%(24/40),有疾病复发组为82.14%(46/56)。结论:AP2S1在BLCA组织中高表达,且与临床分期、病理级别、肿瘤大小以及疾病复发可能密切相关,有望成为一个膀胱癌的治疗靶点。 展开更多
关键词 膀胱尿路上皮癌 ap2S1 免疫组化
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Grain Yield, Quality and 2-Acetyl-1-Pyrroline of Fragrant Rice in Response to Different Planting Seasons in South China 被引量:3
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作者 Leilei Kong Haowen Luo +5 位作者 Zhaowen Mo Shenggang Pan Zhixia Liu Qiang Zhang Song Bai Xiangru Tang 《Phyton-International Journal of Experimental Botany》 SCIE 2020年第3期705-714,共10页
Climate conditions is an important factor affected the fragrant rice growth and development.In order to study the effects of different planting seasons on fragrant rice performance in South China,present study was con... Climate conditions is an important factor affected the fragrant rice growth and development.In order to study the effects of different planting seasons on fragrant rice performance in South China,present study was conducted with three planting seasons(early season(April to July),middle season(June to September)and late season(August to November))and three fragrant rice cultivars,‘Basmati-385’,‘Meixiangzhan-2’and‘Xiangyaxiangzhan’.The results showed that the highest grain yield and grain 2-acetyl-1-pyrroline(2-AP,key component of fragrant rice aroma)content were both recorded in late season treatment while the fragrant rice in middle season treatment produced the lowest grain yield,grain filling percentage,1000-grain weight and gain 2-AP content.The highest contents of precursors(proline,pyrroline-5-carboxylic acid and 1-pyrro-line)which related to 2-AP biosynthesis were recorded in late season treatment compared with early season treatment and middle season treatment.The highest activities of enzymes(proline dehydrogenase,pyrroline-5-carboxylic acid synthetase and ornithine transaminase)which involved in 2-AP biosynthesis were also observed in late season treatment.Moreover,the fragrant rice cultivars in late season possessed the lowest chalk rice rate,chalkiness as well as the highest brown rice rate,head rice and protein content.Thus,the optimal season for fragrant rice production in South China is the late season. 展开更多
关键词 2-acetyl-1-pyrroline climate fragrant rice grain qulity yield formation
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Epoxiconazole Improved Photosynthesis,Yield Formation,Grain Quality and 2-Acetyl-1-Pyrroline Biosynthesis of Fragrant Rice
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作者 LUO Haowen HE Longxin +5 位作者 DU Bin PAN Shenggang MO Zhaowen YANG Shuying ZOU Yingbin TANG Xiangru 《Rice science》 SCIE CSCD 2022年第2期189-196,共8页
Epoxiconazole is a triazole compound.However,the effects of epoxiconazole on crop productivity and quality were rarely reported.In this study,we investigated the effects of epoxiconazole application on yield formation... Epoxiconazole is a triazole compound.However,the effects of epoxiconazole on crop productivity and quality were rarely reported.In this study,we investigated the effects of epoxiconazole application on yield formation,grain quality attributes,and 2-acetyl-1-pyrroline(2-AP)content in fragrant rice.A three-year field experiment was carried out with a fragrant rice variety,Meixiangzhan 2.At the heading stage,0,0.02,0.04,0.08,0.16 and 0.32 g/L epoxiconazole solutions were foliar applied to fragrant rice plants,respectively.The results showed that epoxiconazole application significantly increased grain yield,seed-setting rate and 1000-grain weight.Chlorophyll content and net photosynthetic rate of fragrant rice during the grain-filling stage significantly increased due to epoxiconazole application.Foliar application of epoxiconazole at 0.08 g/L increased grain protein content and decreased both chalky rice rate and chalkiness area ratio of fragrant rice.Epoxiconazole also substantially increased grain 2-AP content by inducing the regulation in contents of related synthetic precursors,including proline,pyrroline-5-carboxylic acid,Δ1-pyrroline and methylglyoxal.Overall,foliar application of epoxiconazole could be used for the improvement in grain yield,grain quality and 2-AP content in fragrant rice production when applied concentration at 0.08-0.32 g/L.Our findings provided the new roles of epoxiconazole in crop production. 展开更多
关键词 EPOXICONAZOLE fragrant rice 2-acetyl-1-pyrroline yield formation grain quality
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Effects of Different Vermicompost Rates on Growth,2-Acetyl-1-Pyrroline,Photosynthesis and Antioxidant Responses of Fragrant Rice(Oryza sativa L.)Seedling
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作者 Shaoyi Ruan Haowen Luo +3 位作者 Xiaoyan Zeng Runhao Wen Feida Wu Xiangru Tang 《Phyton-International Journal of Experimental Botany》 SCIE 2021年第4期1273-1283,共11页
Vermicompost is an organic fertilizer contains multiple nutrient elements.However,the application of vermicompost in fragrant rice production is rarely reported.In order to study the effects of vermicompost applicatio... Vermicompost is an organic fertilizer contains multiple nutrient elements.However,the application of vermicompost in fragrant rice production is rarely reported.In order to study the effects of vermicompost application on fragrant rice(Oryza sativa L.)seedling performances,present study was conducted with two fragrant rice cultivars and four vermicompost rate treatments(0(CK),2.5(Wo1),5.0(Wo2)and 10.0(Wo3)g kg−1).The results showed that vermicompost treatments significantly increased dry weight of fragrant rice seedling by 8.31–32.56%compared with CK.21.10–59.13%higher net photosynthetic rates and 10.66–59.16%higher chlorophyll contents(chlorophyll a,chlorophyll b and total chlorophyll)were recorded in vermicompost treatments than CK.Application of vermicompost also significantly increased 2-acetyl-1-pyrroline(2-AP,the key compound of fragrant rice aroma)content and reduced the transcript level of gene BADH2 which related to 2-AP biosynthesis in fragrant rice seedling.Moreover,compared with CK,vermicompost treatments enhanced activities of superoxide dismutase,peroxidase,catalase by 24.42–28.66%,24.98–25.73%and 22.45–23.57%,respectively.11.54–40.53%lower malonaldehyde contents were recorded in vermicompost treatments in related to CK.In conclusion,vermicompost improved growth,increased 2-AP content and might enhance stress resistant of fragrant rice seedling. 展开更多
关键词 2-acetyl-1-pyrroline fragrant rice gene expression PHOTOSYNTHESIS VERMICOMPOST
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Effects of Exogenousα-Ketoglutaric Acid on 2-Acetyl-1-Pyrroline,Yield Formation and Grain Quality Characters of Aromatic Rice
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作者 Bin Du Qixia Wu +5 位作者 Shuochen Jiang Dingyue Zhang Yue Qiao Yimei Xie Jianlong Xu Jianqiang Zhu 《Phyton-International Journal of Experimental Botany》 SCIE 2021年第2期437-447,共11页
The improvement of grain quality in aromatic rice is very important for farmer to increase their income.Present study was conducted with a two-year field experiment and three aromatic rice cultivars in order to study ... The improvement of grain quality in aromatic rice is very important for farmer to increase their income.Present study was conducted with a two-year field experiment and three aromatic rice cultivars in order to study the effects of exogenousα-ketoglutaric acid on yield formation,grain quality characters and the biosynthesis of 2-acetyl-1-pyrroline(2-AP,key component of aromatic rice’s fragrance)in aromatic rice.At heading stage,0.50 mmol L^(-1)(T1)and 1.00 mmol L^(-1)(T2)α-ketoglutaric acid solutions were overhead sprinkle to aromatic rice plants,respectively while the treatment which was overhead sprinkled with distilled water was set as control(CK).The results showed that 17.34%-33.04%and 21.39%-34.74%higher grain 2-AP contents were recorded in T1 and T2 treatments,respectively.Compared with CK,T1 and T2 treatments significantly reduced the transcript level of gene BADH2 which is related to the 2-AP biosynthesis in aromatic rice.3.86%-7.51%higher grain protein contents and 1.15%-3.37%higher head rice rates were also recorded inα-ketoglutaric acid treatments than CK.Moreover,T1 and T2 treatments remarkably decreased the chalky rice rate,chalkiness and grain amylose content.However,there was no remarkable difference in grain yield and related trails(effective panicle number,grain number per panicle,seed-setting rate and 1000-grain weight)among CK,T1 and T2 treatments.In conclusion,application of exogenousα-ketoglutaric acid enhanced 2-AP biosynthesis and improved grain quality of aromatic rice. 展开更多
关键词 α-ketoglutaric acid aromatic rice CHALKINESS protein 2-acetyl-1-pyrroline
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Enhancement of Ultrasonic Seed Treatment on Yield,Grain Quality Characters,and 2-Acetyl-1-Pyrroline Biosynthesis in Different Fragrant Rice Genotypes
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作者 Rujian Lan Meiyang Duan +4 位作者 Feida Wu Rifang Lai Zhaowen Mo Shenggang Pan Xiangru Tang 《Phyton-International Journal of Experimental Botany》 SCIE 2022年第11期2461-2473,共13页
Fragrant rice is popular for the good grain quality and special aroma.The present study conducted a field experiment to investigate the effects of ultrasonic seed treatment on grain yield,quality characters,physiologi... Fragrant rice is popular for the good grain quality and special aroma.The present study conducted a field experiment to investigate the effects of ultrasonic seed treatment on grain yield,quality characters,physiological properties and aroma biosynthesis of different fragrant rice genotypes.The seeds of three fragrant rice genotypes were exposed to 1 min of ultrasonic vibration and then cultivated in paddy field.The results of present study showed that ultrasonic seed treatment increased grain yield of all fragrant rice genotypes but the responses of yield formation to ultrasonic were varied with different genotypes.Compared with control,ultrasonic seed treatment increased grain 2-acetyl-1-pyrroline(2-AP,the key component of fragrant rice aroma)content by 13.40%–44.88%.Ultrasonic seed treatment also reduced the crude protein contents in grains.The head rice rate,rice length,chalky rice rate,and chalkiness degree were influenced by ultrasonic for one or two fragrant rice genotypes.The activities of peroxidase and superoxide dismutase were also enhanced due to ultrasonic seed treatment.In conclusion,ultrasonic seed treatment increased grain,regulated grain aroma and quality,and improved stress resistance of fragrant rice varieties. 展开更多
关键词 Fragrant rice yield formation crop growth grain quality 2-acetyl-1-pyrroline
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Identification of the E2F1-RAD51AP1 axis as a key factor in MGMT-methylated GBM TMZ resistance
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作者 Junhu Zhou Fei Tong +6 位作者 Jixing Zhao Xiaoteng Cui Yunfei Wang Guangxiu Wang Chunsheng Kang Xiaomin Liu Qixue Wang 《Cancer Biology & Medicine》 SCIE CAS CSCD 2023年第5期385-400,共16页
Objective:Epidermal growth factor receptor variant III(EGFRvIII)is a constitutively-activated mutation of EGFR that contributes to the malignant progression of glioblastoma multiforme(GBM).Temozolomide(TMZ)is a standa... Objective:Epidermal growth factor receptor variant III(EGFRvIII)is a constitutively-activated mutation of EGFR that contributes to the malignant progression of glioblastoma multiforme(GBM).Temozolomide(TMZ)is a standard chemotherapeutic for GBM,but TMZ treatment benefits are compromised by chemoresistance.This study aimed to elucidate the crucial mechanisms leading to EGFRvIII and TMZ resistance.Methods:CRISPR-Cas13a single-cell RNA-seq was performed to thoroughly mine EGFRvIII function in GBM.Western blot,realtime PCR,flow cytometry,and immunofluorescence were used to determine the chemoresistance role of E2F1 and RAD51-associated protein 1(RAD51AP1).Results:Bioinformatic analysis identified E2F1 as the key transcription factor in EGFRvIII-positive living cells.Bulk RNA-seq analysis revealed that E2F1 is a crucial transcription factor under TMZ treatment.Western blot suggested enhanced expression of E2F1 in EGFRvIII-positive and TMZ-treated glioma cells.Knockdown of E2F1 increased sensitivity to TMZ.Venn diagram profiling showed that RAD51AP1 is positively correlated with E2F1,mediates TMZ resistance,and has a potential E2F1 binding site on the promoter.Knockdown of RAD51AP1 enhanced the sensitivity of TMZ;however,overexpression of RAD51AP1 was not sufficient to cause chemotherapy resistance in glioma cells.Furthermore,RAD51AP1 did not impact TMZ sensitivity in GBM cells with high O6-methylguanine-DNA methyltransferase(MGMT)expression.The level of RAD51AP1 expression correlated with the survival rate in MGMT-methylated,but not MGMT-unmethylated TMZ-treated GBM patients.Conclusions:Our results suggest that E2F1 is a key transcription factor in EGFRvIII-positive glioma cells and quickly responds to TMZ treatment.RAD51AP1 was shown to be upregulated by E2F1 for DNA double strand break repair.Targeting RAD51AP1 could facilitate achieving an ideal therapeutic effect in MGMT-methylated GBM cells. 展开更多
关键词 TMZ EGFRVIII E2F1 RAD51ap1 MGMT
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冬小麦miR172响应抗寒的特征分析
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作者 王多佳 王政委 +5 位作者 任志鹏 彭瞰看 田宇 张达 徐庆华 苍晶 《麦类作物学报》 CAS CSCD 北大核心 2023年第10期1304-1310,共7页
东农冬麦1号(Dn1)是我国首个能在黑龙江省高寒地区安全越冬的强抗寒冬小麦品种。为了解tae-miR172在Dn1抗寒中的调控机制,本研究克隆了Dn1中tae-miR172的前体pre-tae-miR172,根据生物信息学分析预测其靶基因为AP2dn1(TraesCS5D02G486600... 东农冬麦1号(Dn1)是我国首个能在黑龙江省高寒地区安全越冬的强抗寒冬小麦品种。为了解tae-miR172在Dn1抗寒中的调控机制,本研究克隆了Dn1中tae-miR172的前体pre-tae-miR172,根据生物信息学分析预测其靶基因为AP2dn1(TraesCS5D02G486600)。通过农杆菌介导的烟草瞬时表达试验,进一步证明tae-miR172靶向AP2dn1;利用qRT-PCR技术对大田越冬期Dn1分蘖节中pre-tae-miR172和Ap2dn1的相对表达量进行分析。结果表明,pre-tae-miR172的表达量随温度降低呈先升后降的变化趋势,而靶基因Ap2dn1的表达量变化趋势则相反,说明tae-miR172负调控Ap2dn1的表达。 展开更多
关键词 冬小麦 tae-miR172 ap2dn1 低温胁迫 表达模式
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沪油15中两个AP2/ERF-B1亚族转录因子的克隆与分析 被引量:16
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作者 庄静 彭日荷 +6 位作者 高峰 付晓燕 朱波 金晓芬 Jian Zhang 熊爱生 姚泉洪 《核农学报》 CAS CSCD 北大核心 2009年第3期435-441,453,共8页
利用油菜EST数据库,以拟南芥ERF-B1亚族转录因子序列为探针,电子克隆拼接得到2个cDNA序列(BnaERFB1-1和BnaERFB1-2),通过PCR和RT-PCR方法分别从甘蓝型油菜沪油15的DNA和cDNA中克隆了上述基因。克隆转录因子BnaERFB1-1-Hy15和BnaERFB1-2-... 利用油菜EST数据库,以拟南芥ERF-B1亚族转录因子序列为探针,电子克隆拼接得到2个cDNA序列(BnaERFB1-1和BnaERFB1-2),通过PCR和RT-PCR方法分别从甘蓝型油菜沪油15的DNA和cDNA中克隆了上述基因。克隆转录因子BnaERFB1-1-Hy15和BnaERFB1-2-Hy15与电子克隆的序列差异很小,分别只有2个和6个氨基酸位点不同,且都没有内含子。从cDNA序列、氨基酸序列的相似性、组成成分、理化性质、疏水性/亲水性、进化树、序列比对、功能域、二级和三级结构等方面进行了分析,结果显示,BnaERFB1-1-Hy15和BnaERFB1-2-Hy15转录因子属于AP2/ERF家族中的ERF-B1亚族,是亲水性蛋白,在蛋白质的三级结构上与拟南芥atERF7相似。EST丰度分析显示,BnaERFB1-1的表达主要集中在种子中,而BnaERFB1-2的表达则主要集中在分生组织中。 展开更多
关键词 ap2/ERF B1亚族 转录因子 克隆 沪油15
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牛乳腺炎无乳链球菌PI-2a菌毛岛屿辅助蛋白AP1、BP、AP2三基因的串联表达及其免疫原性研究 被引量:4
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作者 布日额 王金良 +5 位作者 吴金花 锡林高娃 陈金龙 孙立杰 王华 朝洛蒙 《中国预防兽医学报》 CAS CSCD 北大核心 2018年第2期138-141,共4页
为探究无乳链球菌菌毛岛屿3个亚基AP1、AP2、BP融合蛋白的抗原性,本研究根据该3个亚基基因的主要抗原结构域序列设计并合成引物,通过重叠延伸PCR技术获得上述3个亚基的串联核苷酸片段,并插入pET-30a(+)载体,转化至BL21(DE3)感受态细胞... 为探究无乳链球菌菌毛岛屿3个亚基AP1、AP2、BP融合蛋白的抗原性,本研究根据该3个亚基基因的主要抗原结构域序列设计并合成引物,通过重叠延伸PCR技术获得上述3个亚基的串联核苷酸片段,并插入pET-30a(+)载体,转化至BL21(DE3)感受态细胞后诱导表达,表达蛋白经亲和层析纯化后进行western blot鉴定和小鼠免疫实验。结果表明,AP1-AP2-BP融合抗原具有良好的反应原性和免疫原性。本研究为进一步研究无乳链球菌基于AP1-BP-AP2融合蛋白的致病机制、检测制剂及亚单位疫苗研制提供了实验依据。 展开更多
关键词 无乳链球菌 菌毛岛屿 ap1 BP ap2 串联表达
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桃红四物汤对光老化小鼠皮肤组织中Smad-3、TIMP-1、TIMP-2、AP-1蛋白表达水平的影响 被引量:9
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作者 张宇 张小卿 +1 位作者 吴景东 马贤德 《辽宁中医杂志》 CAS 北大核心 2015年第11期2229-2232,共4页
目的:研究桃红四物汤对光老化小鼠皮肤组织中Smad-3、TIMP-1、TIMP-2、AP-1蛋白含量的影响。方法:将30只小鼠随机分为6组,分别为中药低、中、高剂量组、空白对照组、模型对照组、阳性对照组(VE组)。对模型对照组、阳性对照组(VE组)和中... 目的:研究桃红四物汤对光老化小鼠皮肤组织中Smad-3、TIMP-1、TIMP-2、AP-1蛋白含量的影响。方法:将30只小鼠随机分为6组,分别为中药低、中、高剂量组、空白对照组、模型对照组、阳性对照组(VE组)。对模型对照组、阳性对照组(VE组)和中药低、中、高剂量组小鼠进行皮肤光老化模型制备,空白对照组小鼠正常饲养。在造模前30 min,给空白对照组和模型对照组小鼠灌胃生理盐水,高、中、低剂量组小鼠灌胃相应剂量药液,阳性对照组(VE组)灌胃维生素E滴剂。造模完成后,采用Western-blot方法检测皮肤组织中MMP-1、MMP-3、MMP-12、Smad-2蛋白含量的表达水平。结果:与空白对照组比较,模型对照组小鼠皮肤组织中Smad-3、TIMP-1、TIMP-2蛋白含量的表达水平显著下降,差异具有统计学意义(P<0.05),AP-1蛋白表达水平显著上升(P<0.05);与模型对照组相比,中药低、中、高剂量组小鼠皮肤组织中Smad-3、TIMP-1、TIMP-2蛋白的表达水平显著上升(P<0.05),AP-1蛋白表达水平显著下降(P<0.05)。结论:桃红四物汤对皮肤光老化具有一定的保护作用。 展开更多
关键词 桃红四物汤 光老化 Smad-3 TIMP-1 TIMP-2 ap-1
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p-ERK1/2-AP-1通路在姜黄素抗大鼠糖尿病神经病理性痛中的作用 被引量:8
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作者 黄葱葱 陈果 +3 位作者 吴艳 连庆泉 李军 曹红 《中国病理生理杂志》 CAS CSCD 北大核心 2011年第6期1143-1148,共6页
目的:观察p-ERK1/2-AP-1通路在姜黄素(Cur)抗大鼠糖尿病神经病理性痛(DNP)中的作用。方法:雄性SD大鼠96只,随机分为4组(n=24):正常对照组、DNP组、DNP+溶剂组(DNP+Sol组)和DNP+Cur 100 mg/kg组(DNP+Cur组)。除正常对照组外,其余各组采... 目的:观察p-ERK1/2-AP-1通路在姜黄素(Cur)抗大鼠糖尿病神经病理性痛(DNP)中的作用。方法:雄性SD大鼠96只,随机分为4组(n=24):正常对照组、DNP组、DNP+溶剂组(DNP+Sol组)和DNP+Cur 100 mg/kg组(DNP+Cur组)。除正常对照组外,其余各组采用腹腔注射链唑霉素75 mg/kg的方法制备DNP模型,造模成功后每天1次腹腔注射相应的溶剂或Cur,持续2周。于造模前2 d、造模后14 d、腹腔给药后3、7、14 d时测定机械缩足痛阈(MWT)、热缩足潜伏期(TWL)和非空腹尾静脉血糖值,取脊髓腰膨大及L4/L5背根神经节(DRG),采用免疫组化及Western blotting法测定脊髓背角和DRG p-ERK1/2的表达,电泳迁移率变动分析AP-1的表达。结果:与正常对照组相比,DNP组各时点MWT降低、TWL缩短;血糖值升高;脊髓背角及DRG p-ERK1/2均出现表达上调(P<0.05);脊髓背角AP-1表达上调(P<0.05)。与DNP组相比,DNP+Cur组在给药后7 dMWT回升、TWL延长;在给药后14 d脊髓背角及DRG p-ERK1/2、脊髓背角AP-1表达下调(P<0.05),但并不影响血糖水平。结论:Cur可减轻大鼠糖尿病神经病理性痛,其机制可能与抑制脊髓背角和DRG神经元p-ERK1/2-AP-1通路激活有关。 展开更多
关键词 姜黄素 糖尿病 神经病理性痛 p-ERK1/2-ap-1通路
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B7-1与B7-2对调节人IL-2基因的转录因子NF-κB和AP-1的相同作用 被引量:5
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作者 克晓燕 John Gribben +1 位作者 王晶 王德炳 《中国实验血液学杂志》 CAS CSCD 2002年第6期512-518,共7页
为了解B7共刺激对细胞因子 ,特别是对IL 2mRNA及转录因子NF κB和AP 1的影响 ,探讨B7介导的IL 2调节的分子机制 ,在异基因混合淋巴细胞反应 (MLR)体系中分别或联合加入抗B7 1、抗B7 2单克隆抗体和CTLA 4Ig以阻断B7/CD2 8信号传导 ,通过... 为了解B7共刺激对细胞因子 ,特别是对IL 2mRNA及转录因子NF κB和AP 1的影响 ,探讨B7介导的IL 2调节的分子机制 ,在异基因混合淋巴细胞反应 (MLR)体系中分别或联合加入抗B7 1、抗B7 2单克隆抗体和CTLA 4Ig以阻断B7/CD2 8信号传导 ,通过竞争性PCR定量检测其对IL 2和IL 4mRNA的影响 ,并初步测定IFN γmRNA的改变 ,同时用转染MHCⅡ类分子及联合转染等量B7 1或B7 2的NIH3T3转基因细胞tDR7,tDR7/B7 1和tDR7/B7 2刺激CD2 8+ T细胞 ,通过DNA 蛋白结合实验观察B7对IL 2转录因子NF κB和AP 1的影响。结果表明 :抗B7 2单抗和CTLA 4Ig可明显抑制B7介导的IL 2和IL 4mRNA合成 ,而抗B7 1单抗仅有轻度抑制作用 ,2种或 3种抗体联合应用时抑制作用相加。MLR 1 - 6小时 ,单独tDR7即可诱导NF κB的表达 ,联合转染B7早期对其结合活力无明显影响 ,6小时后tDR7诱导作用减弱 ,B7却可显著延长tDR7的诱导作用至 72小时。tDR7早期同样可诱导AP 1的表达 ,联合转染B7分子在 2 4小时内对其有一定的抑制作用 ,而在反应后期可延长tDR7对AP 1的上调作用 ,B7 1与B7 2间作用未见明显不同。结论 :B7通过减少IL 2mRNA降解和影响基因转录而上调IL 2分泌 ,并可同时影响多种细胞因子分泌 ;在转录水平B7 1与B7 2作用未见明显不同 。 展开更多
关键词 B7-1 B7-2 IL-2基因 NF-кB 转录因子 ap-1 移植免疫
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Adipophilin通过ERK1/2-AP-1途径诱导炎症因子的表达 被引量:3
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作者 袁中华 谭艳美 +5 位作者 陶媛 汪江波 郭东铭 王佐 唐朝克 田国平 《中国病理生理杂志》 CAS CSCD 北大核心 2015年第11期1998-2004,共7页
目的:观察巨噬细胞中脂肪分化相关蛋白(adipophilin)对炎症因子表达的影响,阐明adipophilin促进巨噬细胞炎症因子表达的机制。方法:将已构建成功的adipophilin稳定高、低表达逆转录病毒载体转染入PA317包装细胞,制备adipophilin高和低... 目的:观察巨噬细胞中脂肪分化相关蛋白(adipophilin)对炎症因子表达的影响,阐明adipophilin促进巨噬细胞炎症因子表达的机制。方法:将已构建成功的adipophilin稳定高、低表达逆转录病毒载体转染入PA317包装细胞,制备adipophilin高和低表达的RAW264.7细胞;收集已转染成功的各组细胞上清液,用ELISA方法检测IL-6、TNF-α、MCP-1等炎症因子在细胞培养液中的浓度。Western blot检测各组细胞AP-1、p-AP-1、ERK1/2及p-ERK1/2的蛋白水平;用ERK1/2抑制剂PD98059或AP-1抑制剂curcumin孵育细胞,检测各组细胞中以上指标的变化情况。结果:高表达adipophilin的细胞上清液中炎症因子IL-6、MCP-1和TNF-α浓度明显增高,adipophilin siRNA组细胞中的炎症因子降低;高表达adipophilin的细胞中p-ERK1/2和p-AP-1的蛋白水平增高,adipophilin siRNA组则减少;ERK1/2抑制剂PD98059使AP-1蛋白活性明显下调;给予AP-1抑制剂curcumin后,细胞培养液中的IL-6、MCP-1和TNF-α浓度明显下降。结论:Adipophilin在RAW264.7巨噬细胞中能够促进炎症因子的表达。 展开更多
关键词 ADIPOPHILIN 炎症因子 ERK1/2 ap-1
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大熊猫AP2S1基因cDNA克隆及分析 被引量:1
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作者 杜玉杰 侯怡铃 +1 位作者 田丽 侯万儒 《西南大学学报(自然科学版)》 CAS CSCD 北大核心 2012年第6期50-56,共7页
该研究运用RT-PCR技术,首次从大熊猫Ailuropoda melanoleuca的肌肉组织总RNA中成功克隆了AP2S1基因的编码序列,并对其进行了全面分析.结果表明:大熊猫AP2S1基因编码序列克隆片段全长为512bp,开放阅读框(ORF)为429bp,编码142个氨基酸的... 该研究运用RT-PCR技术,首次从大熊猫Ailuropoda melanoleuca的肌肉组织总RNA中成功克隆了AP2S1基因的编码序列,并对其进行了全面分析.结果表明:大熊猫AP2S1基因编码序列克隆片段全长为512bp,开放阅读框(ORF)为429bp,编码142个氨基酸的蛋白质,该蛋白的相对分子质量为1.7×104,等电点pI为5.82,含有4种类型共5个功能位点,即1个蛋白激酶C磷酸化位点、2个酪蛋白激酶Ⅱ磷酸化位点、1个N-酰基化位点和1个网格蛋白调节素小肽链标签;且该蛋白在哺乳动物网状细胞中的半寿期(half-life)大于30h,不稳定指数(in-stability index)为26.99,属于稳定蛋白.进一步分析发现,大熊猫AP2S1基因与已报道的人、苏门答腊猩猩、牛、褐家鼠和小家鼠5个哺乳动物物种的编码序列及其编码的氨基酸序列具有很高的相似性,表明该基因及其编码蛋白在物种漫长的进化过程中极其保守. 展开更多
关键词 大熊猫 RT-PCR ap2S1 克隆 序列分析
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低盐通过激活ERK和AP-1通路诱导小鼠致密斑细胞系COX-2的表达 被引量:2
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作者 刘冬妍 李学旺 +1 位作者 李航 李雪梅 《基础医学与临床》 CSCD 北大核心 2007年第5期485-489,共5页
目的探讨低盐培养对小鼠致密斑(mouse macula densa derived cells,MMDD1)细胞环氧化酶-2(cyclooxygen-ase,COX-2)表达和p44/42激酶(ERK)、AP-1活性的影响。方法经脂质体转染含AP-1的报告质粒,采用瞬时表达方法检测AP-1转录活性;RT-PCR... 目的探讨低盐培养对小鼠致密斑(mouse macula densa derived cells,MMDD1)细胞环氧化酶-2(cyclooxygen-ase,COX-2)表达和p44/42激酶(ERK)、AP-1活性的影响。方法经脂质体转染含AP-1的报告质粒,采用瞬时表达方法检测AP-1转录活性;RT-PCR检测MMDD1细胞COX-2的表达;免疫印迹方法检测细胞内p-p44/42、C-JUN、C-FOS和COX-2蛋白的表达。用ELISA法检测上清液PGE2的含量。结果低盐(LS)培养促进MMDD1细胞COX-2mRNA和蛋白表达。培养后ERK的磷酸化程度显著上调,180min达到高峰;ERK抑制剂PD-98059降低LS诱导的COX-2表达和PGE2分泌;LS培养促进C-JUN、C-FOS蛋白表达,激活AP-1的转录活性。AP-1抑制剂curcumin(20μmol/L)下调LS诱导的AP-1活性、COX-2mRNA和蛋白表达。结论LS促进MMDD1细胞COX-2的表达,其作用可能与促进ERK的磷酸化、增加AP-1的活性有关。 展开更多
关键词 COX-2 PGE2 ap-1 质粒 ERK
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Cdk2ap1在不同性别鸡胚中的表达 被引量:1
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作者 杨宇 俸艳萍 +4 位作者 龚萍 黄潘 李世军 彭秀丽 龚炎长 《遗传》 CAS CSCD 北大核心 2009年第9期936-940,共5页
为研究通过抑制消减杂交筛选出的cdk2ap1基因在鸡胚胎发育过程中的表达,设计cdk2ap1引物并通过RT-PCR扩增cdk2ap1基因片段,构建cdk2ap1/pGEM-T重组质粒。以构建的重组质粒为模板,使用Sp6和T7RNA聚合酶合成地高辛(dig)标记的正、反义RNA... 为研究通过抑制消减杂交筛选出的cdk2ap1基因在鸡胚胎发育过程中的表达,设计cdk2ap1引物并通过RT-PCR扩增cdk2ap1基因片段,构建cdk2ap1/pGEM-T重组质粒。以构建的重组质粒为模板,使用Sp6和T7RNA聚合酶合成地高辛(dig)标记的正、反义RNA探针,借助胚胎整体原位杂交技术研究了cdk2ap1在雌雄鸡胚中的表达。结果表明cdk2ap1基因在4.0d两性胚胎的脑间质、菱脑、听囊、脊神经管、前肢等部位均有表达,且在雄性鸡胚中的表达量高于雌性;在雄性鸡胚的生殖脊、后肢部位中该基因有表达但在雌性胚胎对应部位却基本没有表达。鉴于该基因在雌雄鸡胚中的差异表达,推测cdk2ap1基因在鸡胚性别分化及性腺发育过程中起一定调节作用。 展开更多
关键词 cdk2ap1 基因克隆 整体原位杂交 鸡胚 性腺发育
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miRNA-330-3p/Ap2m1轴在过表达GATA-4骨髓间充质干细胞外泌体抗心肌细胞凋亡中的作用 被引量:5
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作者 贺继刚 王梓豪 +2 位作者 李敏 谢巧丽 毛富刚 《实用医学杂志》 CAS 北大核心 2020年第20期2763-2768,共6页
目的探讨过表达GATA-4小鼠骨髓间充质干细胞(BMSC)分泌外泌体(BMSC^(GATA-4)exosome)通过miRNA-330-3p/Ap2m1轴减少心肌细胞凋亡,提高心肌梗死心功能。方法建立小鼠心梗模型,共分7组,分别经尾静脉注射BMSC^(miRNA-330-3p-mimic)-exosome... 目的探讨过表达GATA-4小鼠骨髓间充质干细胞(BMSC)分泌外泌体(BMSC^(GATA-4)exosome)通过miRNA-330-3p/Ap2m1轴减少心肌细胞凋亡,提高心肌梗死心功能。方法建立小鼠心梗模型,共分7组,分别经尾静脉注射BMSC^(miRNA-330-3p-mimic)-exosome、BMSC^(miRNA-330-3p-inhibitor)-exosome、BMSC^(空载体)-exosome、BMSCexosome 48 h,同时将心梗未处理组及正常小鼠作为对照组。采用心脏彩超检测各组心功能,RT-PCR检测心梗心肌细胞内miRNA-330-3p的表达,Tunel法检测各组心梗心肌细胞凋亡数量。miRNA-330-3p对应靶基因Ap2.m1、Cnot4蛋白采用Western blot法进行检测。结果BMSC^(miRNA-330-3p-mimic)-exosome组心梗心功能改善最为明显(P<0.05),其心肌细胞内miRNA-330-3p表达量最高(P<0.05)。BMSC^(miRNA-330-3p-mimic)-exosome组心肌细胞的凋亡率最低(P<0.05),BMSC^(miRNA-330-3p-mimic)-exosome组心梗心肌细胞内的Ap2.m1的表达最低(P<0.05)。结论过表达GATA-4小鼠骨髓间充质干细胞外泌体通过miRNA-330-3p/Ap2m1轴抗心肌细胞凋亡。 展开更多
关键词 miRNA-330-3p ap2m1蛋白 GATA-4 外泌体 小鼠骨髓间充质干细胞
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