Aim： To investigate the possible functions of human sperm membrane protein （hSMP-1） in the process of fertilization. Methods： A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was ...Aim： To investigate the possible functions of human sperm membrane protein （hSMP-1） in the process of fertilization. Methods： A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 （anti-rhSMP-1） antibodies. Sperm acrosome reaction and spermzona pellucida （ZP） binding assay were carried out in 10-week-old BALB/c mice. Results： Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 （P 〈 0.05）. Conclusion： The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.展开更多
Lymphotoxin(LT) is a glycoprotein secreted by activated T cell. The expression of LT gene is mainly regulated at the level of transcription. By using human LT DNA as a probe, we carried out a RNA dot blotting test and...Lymphotoxin(LT) is a glycoprotein secreted by activated T cell. The expression of LT gene is mainly regulated at the level of transcription. By using human LT DNA as a probe, we carried out a RNA dot blotting test and found that the longer the time of Jurkat human Tlymphoma cells exposed to the PMA and PHA, the more endogenous LT mRNA could be produced. Results of gel retardation assay showed that the nuclear extract from Jurkat cells treated with PMA and PHA formed different DNA-protein complexes. Changes in complex patterns were observed at various time intervals of PMA and PHA induction. A specific protein-binding site was mapped out to be a 22-bp sequence at the 5’upstream regioll of human LT gene by DNase I footprinting analysis. This region was similar to the sequence recognized by the proteins of NFｋB family The results of fragment competition and homology analysis indicated that the 22-bp sequence contains a ｋB-like motif only which is located at the base pairs -100 to -90 (5’-GGGGGCTTCCC-3’). Thus, the NF-ｋBlike factors were involved in the protein-DNA interaction.Furthermore, there were more than one retarded bands appearing in the gel retardation assay. It suggested that there may be several NF-ｋB-like factors involved in theregulation of LT gene transcription at the same site.展开更多
In recent years,a large number of differentially expressed genes have been identified in human umbilical cord mesenchymal stem cell(hUMSC)transplants for the treatment of ischemic cerebral infarction.These genes are i...In recent years,a large number of differentially expressed genes have been identified in human umbilical cord mesenchymal stem cell(hUMSC)transplants for the treatment of ischemic cerebral infarction.These genes are involved in various biochemical processes,but the role of microRNAs(miRNAs)in this process is still unclear.From the Gene Expression Omnibus(GEO)database,we downloaded two microarray datasets for GSE78731(messenger RNA(mRNA)profile)and GSE97532(miRNA profile).The differentially expressed genes screened were compared between the hUMSC group and the middle cerebral artery occlusion group.Gene ontology enrichment and pathway enrichment analyses were subsequently conducted using the online Database for Annotation,Visualization,and Integrated Discovery.Identified genes were applied to perform weighted gene co-suppression analyses,to establish a weighted co-expression network model.Furthermore,the protein-protein interaction network for differentially expressed genes from turquoise modules was built using Cytoscape(version 3.40)and the most highly correlated subnetwork was extracted from the protein-protein interaction network using the MCODE plugin.The predicted target genes for differentially expressed miRNAs were also identified using the online database starBase v3.0.A total of 3698 differentially expressed genes were identified.Gene ontology analysis demonstrated that differentially expressed genes that are related to hUMSC treatment of ischemic cerebral infarction are involved in endocytosis and inflammatory responses.We identified 12 differentially expressed miRNAs in middle cerebral artery occlusion rats after hUMSC treatment,and these differentially expressed miRNAs were mainly involved in signaling in inflammatory pathways,such as in the regulation of neutrophil migration.In conclusion,we have identified a number of differentially expressed genes and differentially expressed mRNAs,miRNA-mRNAs,and signaling pathways involved in the hUMSC treatment of ischemic cerebral infarction.Bioinformatics and interaction analyses can provide novel clues for further research into hUMSC treatment of ischemic cerebral infarction.展开更多
With the continuing development and improvement of genome-wide techniques, a great number of candidate genes are discovered. How to identify the most likely disease genes among a large number of candidates becomes a f...With the continuing development and improvement of genome-wide techniques, a great number of candidate genes are discovered. How to identify the most likely disease genes among a large number of candidates becomes a fundamental challenge in human health. A common view is that genes related to a specific or similar disease tend to reside in the same neighbourhood of biomolecular networks. Recently, based on such observations,many methods have been developed to tackle this challenge. In this review, we firstly introduce the concept of disease genes, their properties, and available data for identifying them. Then we review the recent computational approaches for prioritizing candidate disease genes based on Protein-Protein Interaction（PPI） networks and investigate their advantages and disadvantages. Furthermore, some pieces of existing software and network resources are summarized. Finally, we discuss key issues in prioritizing candidate disease genes and point out some future research directions.展开更多
文摘Aim： To investigate the possible functions of human sperm membrane protein （hSMP-1） in the process of fertilization. Methods： A 576-bp cDNA fragment of HSD-1 gene coding for the extracellular domain of hSMP-1 was cloned and expressed. The localization of this protein on human and mouse sperm was determined by indirect immunofluorescent staining by using anti-recombinant hSMP-1 （anti-rhSMP-1） antibodies. Sperm acrosome reaction and spermzona pellucida （ZP） binding assay were carried out in 10-week-old BALB/c mice. Results： Recombinant hSMP-1 was successfully cloned and expressed. The expression of the native protein was limited on the acrosome of human and mouse sperm. Treatment of anti-rhSMP-1 antibodies significantly decreased the average number of sperms bound to each egg. Meanwhile, the percentage of acrosome reaction was decreased in comparison to pre-immune control after treatment with anti-rhSMP-1 （P 〈 0.05）. Conclusion： The results suggest that anti-rhSMP-1 antibody inhibited mouse acrosome reaction and sperm-ZP binding.
文摘Lymphotoxin(LT) is a glycoprotein secreted by activated T cell. The expression of LT gene is mainly regulated at the level of transcription. By using human LT DNA as a probe, we carried out a RNA dot blotting test and found that the longer the time of Jurkat human Tlymphoma cells exposed to the PMA and PHA, the more endogenous LT mRNA could be produced. Results of gel retardation assay showed that the nuclear extract from Jurkat cells treated with PMA and PHA formed different DNA-protein complexes. Changes in complex patterns were observed at various time intervals of PMA and PHA induction. A specific protein-binding site was mapped out to be a 22-bp sequence at the 5’upstream regioll of human LT gene by DNase I footprinting analysis. This region was similar to the sequence recognized by the proteins of NFｋB family The results of fragment competition and homology analysis indicated that the 22-bp sequence contains a ｋB-like motif only which is located at the base pairs -100 to -90 (5’-GGGGGCTTCCC-3’). Thus, the NF-ｋBlike factors were involved in the protein-DNA interaction.Furthermore, there were more than one retarded bands appearing in the gel retardation assay. It suggested that there may be several NF-ｋB-like factors involved in theregulation of LT gene transcription at the same site.
文摘目的:观察人胃癌、癌旁组织与胃癌AGS细胞中人音猬因子相互作用蛋白(human hedgehog interacting protein,HHIP)基因启动子区域CpG岛的甲基化水平,探索其与胃癌发生的关系。方法:RT-PCR检测30例人胃癌组织、癌旁组织及AGS细胞中HHIP mRNA的表达,免疫组织化学方法和甲基化特异性PCR(methylation specific PCR,MSP)分别检测胃癌组织和癌旁组织的HHIP表达和HHIP基因启动子区域甲基化状态。AGS细胞予甲基化转移酶抑制剂5-氮杂-2'-脱氧胞苷(5-Aza-2'-deoxycitydine,5-Aza-dc)处理前后,RT-PCR、MSP和硫化测序PCR(bisulfite sequencing PCR,BSP)分别检测AGS细胞中HHIP mRNA表达、启动子区域甲基化水平变化、CpG岛甲基化位点数量的变化;分析HHIP基因启动子区CpG岛甲基化水平变化与HHIP mRNA表达水平变化之间的相关性。结果:胃癌组织中的HHIP mRNA(0.82±0.38 vs 1.60±0.26,P=0.000)和蛋白(0.51±0.03 vs 0.83±0.27,P<0.05)的表达均低于癌旁组织,并且与年龄、性别、TNM分期、分化程度、淋巴结转移均无显著相关性(均P>0.05)。癌旁组织中HHIP基因启动子区甲基化水平显著低于胃癌和AGS细胞[(17.7±3.59)%vs(62.9±6.14)%、(99.7±0.67)%,均P<0.05]。AGS细胞在5-Aza-dc干预后HHIP mRNA表达明显增高(4.68±0.22 vs 0.21±0.12,P<0.01),HHIP基因启动子区甲基化水平明显下降[(10.1±0.21)%vs(90.2±0.67)%,P<0.01],CpG岛甲基化位点明显减少,并且HHIP基因启动子区甲基化水平与mRNA表达呈负相关(r=-0.693,P=0.00)。结论:HHIP基因启动子区CpG岛的高甲基化水平可能通过抑制HHIP基因表达参与胃癌的发生。
基金supported by the National Key Research&Development Program of China,No.2016YFC1301600Program for Jilin University Science and Technology Innovation Team,No.2017TD-12(both to YY)
文摘In recent years,a large number of differentially expressed genes have been identified in human umbilical cord mesenchymal stem cell(hUMSC)transplants for the treatment of ischemic cerebral infarction.These genes are involved in various biochemical processes,but the role of microRNAs(miRNAs)in this process is still unclear.From the Gene Expression Omnibus(GEO)database,we downloaded two microarray datasets for GSE78731(messenger RNA(mRNA)profile)and GSE97532(miRNA profile).The differentially expressed genes screened were compared between the hUMSC group and the middle cerebral artery occlusion group.Gene ontology enrichment and pathway enrichment analyses were subsequently conducted using the online Database for Annotation,Visualization,and Integrated Discovery.Identified genes were applied to perform weighted gene co-suppression analyses,to establish a weighted co-expression network model.Furthermore,the protein-protein interaction network for differentially expressed genes from turquoise modules was built using Cytoscape(version 3.40)and the most highly correlated subnetwork was extracted from the protein-protein interaction network using the MCODE plugin.The predicted target genes for differentially expressed miRNAs were also identified using the online database starBase v3.0.A total of 3698 differentially expressed genes were identified.Gene ontology analysis demonstrated that differentially expressed genes that are related to hUMSC treatment of ischemic cerebral infarction are involved in endocytosis and inflammatory responses.We identified 12 differentially expressed miRNAs in middle cerebral artery occlusion rats after hUMSC treatment,and these differentially expressed miRNAs were mainly involved in signaling in inflammatory pathways,such as in the regulation of neutrophil migration.In conclusion,we have identified a number of differentially expressed genes and differentially expressed mRNAs,miRNA-mRNAs,and signaling pathways involved in the hUMSC treatment of ischemic cerebral infarction.Bioinformatics and interaction analyses can provide novel clues for further research into hUMSC treatment of ischemic cerebral infarction.
文摘With the continuing development and improvement of genome-wide techniques, a great number of candidate genes are discovered. How to identify the most likely disease genes among a large number of candidates becomes a fundamental challenge in human health. A common view is that genes related to a specific or similar disease tend to reside in the same neighbourhood of biomolecular networks. Recently, based on such observations,many methods have been developed to tackle this challenge. In this review, we firstly introduce the concept of disease genes, their properties, and available data for identifying them. Then we review the recent computational approaches for prioritizing candidate disease genes based on Protein-Protein Interaction（PPI） networks and investigate their advantages and disadvantages. Furthermore, some pieces of existing software and network resources are summarized. Finally, we discuss key issues in prioritizing candidate disease genes and point out some future research directions.
