OBJECTIVE o facilitate the basic acquaintance with the bioactive lycodine-type alkaloids biosynthetic pathways,we conducted the transcriptome analysis of L.casuarinoides by illumina sequencing.METHODS The plant of L.c...OBJECTIVE o facilitate the basic acquaintance with the bioactive lycodine-type alkaloids biosynthetic pathways,we conducted the transcriptome analysis of L.casuarinoides by illumina sequencing.METHODS The plant of L.casuarinoides was collected and was subjected to RNA isolation,cDNA library construction,and illumina sequencing before bioinformatics analysis.After sequencing,the clean reads were obtained for de novo assembly by using Trinity software,and then further processed with TGICL sequencing clustering software to generate unigenes,The unigenes are aligned by Blast X alignment to six public protein database.In addition,all unigenes are functionally annotated by GO,KEGG and characterized putative genes involved in lycopodium alkaloids biosynthesis.RESULTS In total,124,524 high-quality unigenes were obtained with an average sequence length of 601 bp.Among the L.casuarinoides transcripts,61,304 showed significant similarity(E-value<1 e-5) to the known proteins in the public database.Among the total 124 524 unigenes,47,538 open reading frame(ORFs) were predicted.Based on the bioinformatics analysis,all possible enzymes involved in the Lycodine-type alkaloids biosynthetic pathway of L.casuarinoides were identified,including primary amine oxidase(PAO),and Malonly-CoA decarboxylase.In addition,a total of 64 putative cytochrome P450(CYP450) and 827 transcription factors were selected as the candidates of Lycodine-type alka.loids modifiers.Furthermore,a total of 13 352 simple sequence repeats(SSRs) were identified from the 124,524 unigenes,of which dinucleotide motifs AG/CT(50.1%),were the most abundant.CONCLU.SION This transcriptome analysis of L.casuarinoides,provides many valuable candidate genes involv.ing in the biosynthesis of novel secondary metabolites but also lays the foundation for genetic diversity analysis via SSRs markers in L.casuarinoides.展开更多
Spinocerebellar ataxia type 3/Machado-Joseph disease （SCA3/MJD） is a progressive, currently untreatable and ultimately fatal ataxic disorder that belongs to the group of neurological disorders known as CAG-repeat or...Spinocerebellar ataxia type 3/Machado-Joseph disease （SCA3/MJD） is a progressive, currently untreatable and ultimately fatal ataxic disorder that belongs to the group of neurological disorders known as CAG-repeat or polyglutamine diseases. Here, we present the first prenatal diagnosis of SCA3/MJD in China's Mainland in a woman who was known to carry an expanded CAG-trinucleotide repeat in the MJD1 gene. After evaluating motivation and psychological tolerance of the couple, amniocentesis was performed after 14 weeks of gestation. Polymerase chain reactions followed by T-vector cloning and direct sequencing were employed to evaluate the CAG-repeat number of the fetal MJD1 gene. We identified a truncated CAG expansion of 78 repeats in the MJD1 gene of the fetus compared with 81 repeats in his mother.展开更多
AIM:To characterize the regeneration-associated stem cell-related phenotype of hepatocyte-derived growth factor receptor(HGFR)-expressing cells in active ulcerative colitis(UC).METHODS:On the whole 38 peripheral blood...AIM:To characterize the regeneration-associated stem cell-related phenotype of hepatocyte-derived growth factor receptor(HGFR)-expressing cells in active ulcerative colitis(UC).METHODS:On the whole 38 peripheral blood samples and 38 colonic biopsy samples from 18 patients with histologically proven active UC and 20 healthy control subjects were collected.After preparing tissue microarrays and blood smears HGFR,caudal type homeobox 2(CDX2),prominin-1(CD133) and Musashi-1conventional and double fluorescent immunolabelings were performed.Immunostained samples were digitalized using high-resolution Mirax Desk instrument,and analyzed with the Mirax TMA Module software.For semiquantitative counting of immunopositive lamina propria(LP) cells 5 fields of view were counted at magnification x 200 in each sample core,then mean ± SD were determined.In case of peripheral blood smears,30 fields of view with 100 μm diameter were evaluated in every sample and the number of immunopositive cells(mean ± SD) was determined.Using 337 nm UVA Laser MicroDissection system at least 5000 subepithelial cells from the lamina propria were collected.Gene expression analysis of HGFR,CDX2,CD133,leucine-rich repeat-containing G-protein coupled receptor 5(Lgr5),Musashi-1 and cytokeratin20(CK20) were performed in both laser-microdisscted samples and blood samples by using real time reverse transcription polymerase chain reaction(RT-PCR).RESULTS:By performing conventional and double fluorescent immunolabelings confirmed by RT-PCR,higher number of HGFR(blood:6.7 ± 1.22 vs 38.5 ±3.18;LP:2.25 ± 0.85 vs 9.22 ± 0.65;P < 0.05),CDX2(blood:0 vs 0.94 ± 0.64;LP:0.75 ± 0.55 vs 2.11± 0.75;P < 0.05),CD133(blood:1.1 ± 0.72 vs 8.3± 1.08;LP:11.1 ± 0.85 vs 26.28 ± 1.71;P < 0.05)and Musashi-1(blood and LP:0 vs scattered) positive cells were detected in blood and lamina propria of UC samples as compared to controls.HGFR/CDX2(blood:0 vs 1± 0.59;LP:0.8 ± 0.69 vs 2.06 ± 0.72,P < 0.05)and Musashi-1/CDX2(blood and LP:0 vs scattered) coexpressions were found in blood and lamina propria of UC samples.HGFR/CD133 and CD133/CDX2 coexpressions appeared only in UC lamina propria samples.CDX2,Lgr5 and Musashi-1 expressions in UC blood samples were not accompanied by CK20 mRNA expression.CONCLUSION:In active UC,a portion of circulating HGFR-expressing cells are committed to the epithelial lineage,and may participate in mucosal regeneration by undergoing mesenchymal-to-epithelial transition.展开更多
目的探讨血浆金属肽酶含血小板反应蛋白13(a disintegrin-like and metalloprotease with thrombospondin type 1 repeats-13,ADAMTS13)水平对心力衰竭合并2型糖尿病(type 2 diabetes mellitus,T2DM)患者的预后价值。方法收集2019年1月...目的探讨血浆金属肽酶含血小板反应蛋白13(a disintegrin-like and metalloprotease with thrombospondin type 1 repeats-13,ADAMTS13)水平对心力衰竭合并2型糖尿病(type 2 diabetes mellitus,T2DM)患者的预后价值。方法收集2019年1月至2020年12月在榆林市第二医院住院的急性失代偿性心力衰竭成年患者临床资料,共纳入278例。采用ELISA试剂盒测定ADAMTS13水平,并基于ADAMTS13三分位数将患者分为T1<9.9μg/L组(n=92),T2≥10.0~18.5μg/L组(n=93)和T3≥18.5μg/L组(n=93)。主要结局定义为全因死亡率或心力衰竭再住院的复合终点。次要结局包括全因死亡率和心力衰竭再住院。结果与ADAMTS13最高三分位数的患者相比,ADAMTS13最低三分位数的患者中T2DM例数、身体质量指数、尿酸、空腹血糖、糖化血红蛋白(glycated hemoglobin,HbA1c)和超敏C反应蛋白(high sensitivity C-reactive protein,hs-CRP)水平均显著增加(P<0.05)。T2DM患者的主要结局、全因死亡和心力衰竭住院的绝对风险高于非T2DM患者(χ^(2)=12.028、8.725、9.023,均P=0.000)。不同ADAMTS13水平患者心力衰竭率的差异有统计学意义(χ^(2)=9.384,P=0.002)。ADAMTS13每增加一倍与合并T2DM的HF患者的主要结局(HR=0.69,95%CI=0.61~0.79)、全因死亡率(HR=0.59,95%CI=0.47-0.74)和心衰再住院(HR=0.75,95%CI=0.65~0.87)下降相关。结论ADAMTS13是HF危险分层的一个独立的生物标志物,特别是在合并T2DM的患者中。展开更多
粘虫 Mythimna separata（Walker）是一种迁飞性害虫，严重危害玉米、水稻、小麦等粮食作物。 SSR 是指以1～6个核苷酸为基本重复单位的串联重复 DNA 序列。 SSR 位点的信息分析为粘虫扩散、迁飞和交配等行为分子机制的研究以及粘虫的...粘虫 Mythimna separata（Walker）是一种迁飞性害虫，严重危害玉米、水稻、小麦等粮食作物。 SSR 是指以1～6个核苷酸为基本重复单位的串联重复 DNA 序列。 SSR 位点的信息分析为粘虫扩散、迁飞和交配等行为分子机制的研究以及粘虫的综合防治奠定理论基础。本研究基于高通量测序获得的粘虫转录组数据，利用软件 msatcom‐mander 发掘粘虫 SSR 位点。结果从20776条转录组 Unigenes 中共搜索出400个 SSR ，分布于372条 Unigenes 中。在粘虫转录组 SSR 中，三核苷酸重复的数量最为丰富，有271个；其次是二核苷酸和单核苷酸重复，分别是70个和49个；四至六核苷酸重复的数量都很少，共10个。粘虫转录组 SSR 共包含24种重复基元，其中 CCG／CGG 是优势重复基元类型，有69个；其次是 AAG／CTT ，有57个。 CG／CG 有18个，在二核苷酸重复基元中所占的比例达到25.7％。此研究发掘到的 SSR 位点将为粘虫遗传图谱的构建、遗传多样性分析、亲缘关系分析等提供丰富的分子标记。展开更多
基金supported by Jiangxi′s Major International Science and Technology Cooperation Projects(20151BDH80020)
文摘OBJECTIVE o facilitate the basic acquaintance with the bioactive lycodine-type alkaloids biosynthetic pathways,we conducted the transcriptome analysis of L.casuarinoides by illumina sequencing.METHODS The plant of L.casuarinoides was collected and was subjected to RNA isolation,cDNA library construction,and illumina sequencing before bioinformatics analysis.After sequencing,the clean reads were obtained for de novo assembly by using Trinity software,and then further processed with TGICL sequencing clustering software to generate unigenes,The unigenes are aligned by Blast X alignment to six public protein database.In addition,all unigenes are functionally annotated by GO,KEGG and characterized putative genes involved in lycopodium alkaloids biosynthesis.RESULTS In total,124,524 high-quality unigenes were obtained with an average sequence length of 601 bp.Among the L.casuarinoides transcripts,61,304 showed significant similarity(E-value<1 e-5) to the known proteins in the public database.Among the total 124 524 unigenes,47,538 open reading frame(ORFs) were predicted.Based on the bioinformatics analysis,all possible enzymes involved in the Lycodine-type alkaloids biosynthetic pathway of L.casuarinoides were identified,including primary amine oxidase(PAO),and Malonly-CoA decarboxylase.In addition,a total of 64 putative cytochrome P450(CYP450) and 827 transcription factors were selected as the candidates of Lycodine-type alka.loids modifiers.Furthermore,a total of 13 352 simple sequence repeats(SSRs) were identified from the 124,524 unigenes,of which dinucleotide motifs AG/CT(50.1%),were the most abundant.CONCLU.SION This transcriptome analysis of L.casuarinoides,provides many valuable candidate genes involv.ing in the biosynthesis of novel secondary metabolites but also lays the foundation for genetic diversity analysis via SSRs markers in L.casuarinoides.
基金grants from the National Science and Technology Pillar Program in the Eleventh Five-year Plan Period, No. 2006BAI05A07the Major State Basic Research Development Program of China (973 Program), No. 2006cb500700+5 种基金the National Key Technologies Research and Development Program of China, No. 2004BA720A03the National Natural Science Foundation of China, No. 30871354, 30710303061 and 30470619the Key Project in the Natural Science Foundation of Hunan Province, No. 08JJ3048the Natural Science Foundation of Hunan Province, No. 11JJ5071the Science and Technology Planning Project of Hunan Province, No. 2009SK3172the Graduate Degree Thesis Innovation Foundation of Central South University, No. 2008yb030
文摘Spinocerebellar ataxia type 3/Machado-Joseph disease （SCA3/MJD） is a progressive, currently untreatable and ultimately fatal ataxic disorder that belongs to the group of neurological disorders known as CAG-repeat or polyglutamine diseases. Here, we present the first prenatal diagnosis of SCA3/MJD in China's Mainland in a woman who was known to carry an expanded CAG-trinucleotide repeat in the MJD1 gene. After evaluating motivation and psychological tolerance of the couple, amniocentesis was performed after 14 weeks of gestation. Polymerase chain reactions followed by T-vector cloning and direct sequencing were employed to evaluate the CAG-repeat number of the fetal MJD1 gene. We identified a truncated CAG expansion of 78 repeats in the MJD1 gene of the fetus compared with 81 repeats in his mother.
基金Cell Analysis Laboratory, 2nd Department of Internal Medicine, and the 1st Department of Pathology and Experimental Oncology, Semmelweis University for their technical support
文摘AIM:To characterize the regeneration-associated stem cell-related phenotype of hepatocyte-derived growth factor receptor(HGFR)-expressing cells in active ulcerative colitis(UC).METHODS:On the whole 38 peripheral blood samples and 38 colonic biopsy samples from 18 patients with histologically proven active UC and 20 healthy control subjects were collected.After preparing tissue microarrays and blood smears HGFR,caudal type homeobox 2(CDX2),prominin-1(CD133) and Musashi-1conventional and double fluorescent immunolabelings were performed.Immunostained samples were digitalized using high-resolution Mirax Desk instrument,and analyzed with the Mirax TMA Module software.For semiquantitative counting of immunopositive lamina propria(LP) cells 5 fields of view were counted at magnification x 200 in each sample core,then mean ± SD were determined.In case of peripheral blood smears,30 fields of view with 100 μm diameter were evaluated in every sample and the number of immunopositive cells(mean ± SD) was determined.Using 337 nm UVA Laser MicroDissection system at least 5000 subepithelial cells from the lamina propria were collected.Gene expression analysis of HGFR,CDX2,CD133,leucine-rich repeat-containing G-protein coupled receptor 5(Lgr5),Musashi-1 and cytokeratin20(CK20) were performed in both laser-microdisscted samples and blood samples by using real time reverse transcription polymerase chain reaction(RT-PCR).RESULTS:By performing conventional and double fluorescent immunolabelings confirmed by RT-PCR,higher number of HGFR(blood:6.7 ± 1.22 vs 38.5 ±3.18;LP:2.25 ± 0.85 vs 9.22 ± 0.65;P < 0.05),CDX2(blood:0 vs 0.94 ± 0.64;LP:0.75 ± 0.55 vs 2.11± 0.75;P < 0.05),CD133(blood:1.1 ± 0.72 vs 8.3± 1.08;LP:11.1 ± 0.85 vs 26.28 ± 1.71;P < 0.05)and Musashi-1(blood and LP:0 vs scattered) positive cells were detected in blood and lamina propria of UC samples as compared to controls.HGFR/CDX2(blood:0 vs 1± 0.59;LP:0.8 ± 0.69 vs 2.06 ± 0.72,P < 0.05)and Musashi-1/CDX2(blood and LP:0 vs scattered) coexpressions were found in blood and lamina propria of UC samples.HGFR/CD133 and CD133/CDX2 coexpressions appeared only in UC lamina propria samples.CDX2,Lgr5 and Musashi-1 expressions in UC blood samples were not accompanied by CK20 mRNA expression.CONCLUSION:In active UC,a portion of circulating HGFR-expressing cells are committed to the epithelial lineage,and may participate in mucosal regeneration by undergoing mesenchymal-to-epithelial transition.
文摘目的:探讨细胞分泌蛋白1(C-type lectin repeat protein-1,CTRP1)、细胞分泌蛋白3(C-type lectin repeat protein-3,CTRP3)水平变化在评估阿替普酶+替罗非班治疗超急性期缺血性脑卒中的临床价值。方法:研究本院2021年06月至2024年01月纳入的92例超急性期缺血性脑卒中患者随机分为观察组、对照组各46例。对照组实施阿替普酶静脉溶栓治疗,观察组给予阿替普酶静脉溶栓后序贯替罗非班治疗。比较两组治疗效果,Barthel评分和美国卫生部卒中量表(National Institute of Health Stroke Scale Score,NIHSS)评分,CTRP1和CTRP3水平,血脂水平,不良事件发生率情况。结果:治疗总有效率观察组高于对照组(P<0.05)。治疗后两组Barthel评分和NIHSS评分均改善,与对照组相比观察组更优(P<0.05)。治疗后12 h、24 h两组CTRP1和CTRP3水平均依次降低,与对照组相比,观察组CTRP1和CTRP3水平更低,(P<0.05)。治疗后两组血脂水平均降低,与对照组相比,观察组各血脂水平更低(P<0.05)。观察组总不良事件发生率低于对照组(P<0.05)。结论:阿替普酶+替罗非班在超急性期缺血性脑卒中患者治疗中可有效改善患者的日常生活能力及神经功能缺损,降低患者的血脂水平,CTRP1、CTRP3水平逐渐降低,不良事件发生率较低,临床疗效显著。
文摘粘虫 Mythimna separata（Walker）是一种迁飞性害虫，严重危害玉米、水稻、小麦等粮食作物。 SSR 是指以1～6个核苷酸为基本重复单位的串联重复 DNA 序列。 SSR 位点的信息分析为粘虫扩散、迁飞和交配等行为分子机制的研究以及粘虫的综合防治奠定理论基础。本研究基于高通量测序获得的粘虫转录组数据，利用软件 msatcom‐mander 发掘粘虫 SSR 位点。结果从20776条转录组 Unigenes 中共搜索出400个 SSR ，分布于372条 Unigenes 中。在粘虫转录组 SSR 中，三核苷酸重复的数量最为丰富，有271个；其次是二核苷酸和单核苷酸重复，分别是70个和49个；四至六核苷酸重复的数量都很少，共10个。粘虫转录组 SSR 共包含24种重复基元，其中 CCG／CGG 是优势重复基元类型，有69个；其次是 AAG／CTT ，有57个。 CG／CG 有18个，在二核苷酸重复基元中所占的比例达到25.7％。此研究发掘到的 SSR 位点将为粘虫遗传图谱的构建、遗传多样性分析、亲缘关系分析等提供丰富的分子标记。
