The transcriptional factor GATA-6 gene produces two translational isoforms from a single mRNA through ribosomal leaky scanning. L-type GATA-6 has an extension of 146 amino acid residues at its amino terminus. In the e...The transcriptional factor GATA-6 gene produces two translational isoforms from a single mRNA through ribosomal leaky scanning. L-type GATA-6 has an extension of 146 amino acid residues at its amino terminus. In the extension, there is a unique PEST sequence (Glu31-Cys46), which is composed of an amino terminal Pro-rich segment and a carboxyl terminal Ser-cluster. Substitution of either half of the PEST sequence with Ala residues by cassette mutagenesis reduced the apparent molecular size of L-type GATA-6 on SDS-polyacrylamide gel-electrophoresis. However, the effect of substitution of the Pro-rich segment was much more significant;the mobility increase of the Pro-rich segment on the gel was 13% while that of the Ser-cluster was 8%. Substitution of each amino acid residue demonstrated that the effect of Pro substitution is greater than that of the Ser and Thr residues. Such increased mobility of L-type GATA-6 in the presence of a detergent may apparently correlate with the decrease in transcription activity in vivo as determined by means of luciferase reporter gene assay. The activity of ΔAla (with Ala residues instead of the PEST sequence) was reduced to one fifth of that of ΔA (with the PEST sequence). These results suggest that the PEST sequence of L-type GATA-6 does not function as a constitutive protein degradation signal, but rather plays structural and functional roles in the activation of gene expression on the GATA responsive promoter.展开更多
Heart failure (HF) incidence could cause further complications to other body organs, which might sometimes be fatal, and is accompanied by various biochemical alterations i.e. enzymatic changes. The objective of this ...Heart failure (HF) incidence could cause further complications to other body organs, which might sometimes be fatal, and is accompanied by various biochemical alterations i.e. enzymatic changes. The objective of this study was to measure the activity of gamma glutamyl transferase (GGT) as an early diagnostic indicator for HF patients;and to isolate the iso-enzymes for the purpose of finding the Michaelis-Menten constant (Km) and the maximum velocity Vmax of each iso-enzymes which enable follow up the development of HF disease. Samples of blood sera were collected from 120 patients of both genders (70 males and 50 female, aged 30 - 38 years old). Partial purification of iso-enzyme GGT was performed by precipitation, gel filtration, and ion exchange of the two iso-enzyme (I and II). The purity of the enzyme was confirmed by using Sodium dodecyl sulfate-polyacryl amide gel electrophoresis (SDS-PAGE) into two clear bands. The results were compared with other 80 samples of healthy volunteers whose ages ranged between 25 - 78 years old, used as control. There has been a significant increase (p ≤ 0.01) in the activity of the enzyme GGT in the heart failure patient (66.9 ± 1.7 IU/L) in comparison with control (12.07 ± 0.60 IU/L). It is concluded that measurements of the iso-enzyme GGT could well benefit as a clear indicator criteria in prognosis of heart failure.展开更多
A specific reagent DACM [N-( 7-Dimethylamino-4-methyl-3-coumarinyl) maleimide] is used to study the -SH groups in lens proteins of normal and galactose cataractous rats. DACM when reacts readily with -SH groups form s...A specific reagent DACM [N-( 7-Dimethylamino-4-methyl-3-coumarinyl) maleimide] is used to study the -SH groups in lens proteins of normal and galactose cataractous rats. DACM when reacts readily with -SH groups form strong fluorescent adducts. The two -dimensional electrophoresis with DACM pre-labeled proteins is a simple and sensitive method for detecting -SH groups of protein subunit. In the present study, based on IEF/SDS-PAGE electrophoretically characterized soluble crystallins, describes specific ...展开更多
为了解不同性状窖泥细菌群落结构及酸酯代谢的差异,分别选取新窖泥、趋老熟窖泥和老熟窖泥,对其细菌16S r DNA的V3区进行变性梯度凝胶电泳分析和同源性比较,并结合窖泥主要有机酸及有机酸酯含量进行了典型相关分析。结果表明,老熟窖泥...为了解不同性状窖泥细菌群落结构及酸酯代谢的差异,分别选取新窖泥、趋老熟窖泥和老熟窖泥,对其细菌16S r DNA的V3区进行变性梯度凝胶电泳分析和同源性比较,并结合窖泥主要有机酸及有机酸酯含量进行了典型相关分析。结果表明,老熟窖泥的细菌多样性指数及均匀度指数高于新窖泥和趋老熟窖泥,得到的39个优势条带,进行细菌DNA测序可分为14类;Clostridium XIVa、Aminobacterium均只在老熟窖泥中检测到;新窖泥和趋老熟窖泥与Lactococcus、Lactobacillus、乳酸、乳酸乙酯含量正相关,老熟窖泥与Clostridiales、己酸、己酸乙酯和丁酸含量正相关。展开更多
目的利用16S r DNA PCR及变性梯度凝胶电泳（DGGE）技术,探讨生后经验性应用抗生素对早产儿早期肠道菌群的影响。方法以2014年1月至2015年1月在重庆医科大学附属儿童医院住院治疗的早产儿为研究对象,使用哌拉西林-他唑巴坦〉7 d为哌拉...目的利用16S r DNA PCR及变性梯度凝胶电泳（DGGE）技术,探讨生后经验性应用抗生素对早产儿早期肠道菌群的影响。方法以2014年1月至2015年1月在重庆医科大学附属儿童医院住院治疗的早产儿为研究对象,使用哌拉西林-他唑巴坦〉7 d为哌拉西林组,7 d内未使用抗生素为对照组。收集胎粪（〈12 h）和3、5、7日龄（d3、d5、d7）的粪便标本,采用PCR-DGGE方法对肠道菌群的变化行动态观察,同时行克隆测序分析菌群组成的变化。结果 24例早产儿的96份粪便标本进入分析,哌拉西林组和对照组各12例,两组基线具可比性。24份胎粪提取细菌DNA后行PCR扩增失败,未行DGGE分析。1哌拉西林组d3、d5、d7时点的Shannon指数中位数（P25-P75）分别为1.64（1.16-1.92）、1.97（1.69-2.20）和1.22（0.69-2.10）;对照组分别为1.39（0.94-1.94）、2.24（2.07-2.49）和2.38（2.07-2.61）,哌拉西林组d7时点的Shannon指数显著低于对照组（P〈0.05）。2随日龄增加,对照组Shannon指数逐渐升高;哌拉西林组Shannon指数呈降低趋势。3d7时点哌拉西林组和对照组均以克雷伯菌属占绝对优势（35.5%和42.4%）,且乳酸杆菌属检出率均较低（1.6%和0.8%）。哌拉西林组较对照组肠球菌属（21.0%vs 7.2%）及链球菌属（25.8%vs 4.0%）比例明显增加,肠杆菌比例明显降低（3.2%vs 12.8%）。结论早产儿菌群结构简单,生后抗生素应用会降低肠道菌群多样性。展开更多
文摘The transcriptional factor GATA-6 gene produces two translational isoforms from a single mRNA through ribosomal leaky scanning. L-type GATA-6 has an extension of 146 amino acid residues at its amino terminus. In the extension, there is a unique PEST sequence (Glu31-Cys46), which is composed of an amino terminal Pro-rich segment and a carboxyl terminal Ser-cluster. Substitution of either half of the PEST sequence with Ala residues by cassette mutagenesis reduced the apparent molecular size of L-type GATA-6 on SDS-polyacrylamide gel-electrophoresis. However, the effect of substitution of the Pro-rich segment was much more significant;the mobility increase of the Pro-rich segment on the gel was 13% while that of the Ser-cluster was 8%. Substitution of each amino acid residue demonstrated that the effect of Pro substitution is greater than that of the Ser and Thr residues. Such increased mobility of L-type GATA-6 in the presence of a detergent may apparently correlate with the decrease in transcription activity in vivo as determined by means of luciferase reporter gene assay. The activity of ΔAla (with Ala residues instead of the PEST sequence) was reduced to one fifth of that of ΔA (with the PEST sequence). These results suggest that the PEST sequence of L-type GATA-6 does not function as a constitutive protein degradation signal, but rather plays structural and functional roles in the activation of gene expression on the GATA responsive promoter.
文摘Heart failure (HF) incidence could cause further complications to other body organs, which might sometimes be fatal, and is accompanied by various biochemical alterations i.e. enzymatic changes. The objective of this study was to measure the activity of gamma glutamyl transferase (GGT) as an early diagnostic indicator for HF patients;and to isolate the iso-enzymes for the purpose of finding the Michaelis-Menten constant (Km) and the maximum velocity Vmax of each iso-enzymes which enable follow up the development of HF disease. Samples of blood sera were collected from 120 patients of both genders (70 males and 50 female, aged 30 - 38 years old). Partial purification of iso-enzyme GGT was performed by precipitation, gel filtration, and ion exchange of the two iso-enzyme (I and II). The purity of the enzyme was confirmed by using Sodium dodecyl sulfate-polyacryl amide gel electrophoresis (SDS-PAGE) into two clear bands. The results were compared with other 80 samples of healthy volunteers whose ages ranged between 25 - 78 years old, used as control. There has been a significant increase (p ≤ 0.01) in the activity of the enzyme GGT in the heart failure patient (66.9 ± 1.7 IU/L) in comparison with control (12.07 ± 0.60 IU/L). It is concluded that measurements of the iso-enzyme GGT could well benefit as a clear indicator criteria in prognosis of heart failure.
文摘A specific reagent DACM [N-( 7-Dimethylamino-4-methyl-3-coumarinyl) maleimide] is used to study the -SH groups in lens proteins of normal and galactose cataractous rats. DACM when reacts readily with -SH groups form strong fluorescent adducts. The two -dimensional electrophoresis with DACM pre-labeled proteins is a simple and sensitive method for detecting -SH groups of protein subunit. In the present study, based on IEF/SDS-PAGE electrophoretically characterized soluble crystallins, describes specific ...
文摘为了解不同性状窖泥细菌群落结构及酸酯代谢的差异,分别选取新窖泥、趋老熟窖泥和老熟窖泥,对其细菌16S r DNA的V3区进行变性梯度凝胶电泳分析和同源性比较,并结合窖泥主要有机酸及有机酸酯含量进行了典型相关分析。结果表明,老熟窖泥的细菌多样性指数及均匀度指数高于新窖泥和趋老熟窖泥,得到的39个优势条带,进行细菌DNA测序可分为14类;Clostridium XIVa、Aminobacterium均只在老熟窖泥中检测到;新窖泥和趋老熟窖泥与Lactococcus、Lactobacillus、乳酸、乳酸乙酯含量正相关,老熟窖泥与Clostridiales、己酸、己酸乙酯和丁酸含量正相关。
