Parthenogenetic embryonic stem cells have pluripotent differentiation potentials, akin to fertilized embryo-derived embryonic stem cells. The aim of this study was to compare the neuronal differentiation potential of ...Parthenogenetic embryonic stem cells have pluripotent differentiation potentials, akin to fertilized embryo-derived embryonic stem cells. The aim of this study was to compare the neuronal differentiation potential of parthenogenetic and fertilized embryo-derived embryonic stem cells. Before differentiation, karyotype analysis was performed, with normal karyotypes detected in both parthenogenetic and fertilized embryo-derived embryonic stem cells. Sex chromosomes were identified as XX. Immunocytochemistry and quantitative real-time PCR detected high expression of the pluripotent gene, Oct4, at both the mRNA and protein levels, indicating pluripotent differentiation potential of the two embryonic stem cell subtypes. Embryonic stern cells were induced with retinoic acid to form embryoid bodies, and then dispersed into single cells. Single cells were differentiated in N2 differentiation medium for 9 days. Immunocytochemistry showed parthenogenetic and fertilized embryo-derived embryonic stem cells both express the neuronal cell markers nestin, ~lll-tubulin and myelin basic protein. Quantitative real-time PCR found expression of neuregenesis related genes （Sox-1, Nestin, GABA, Pax6, Zic5 and Pitxl） in both types of embryonic stem cells, and Oct4 expression was significantly decreased. Nestin and Pax6 expression in parthenogenetic embryonic stem cells was significantly higher than that in fertilized embryo-derived embryonic stem cells. Thus, our experimental findings indicate that parthenogenetic embryonic stem cells have stronger neuronal differentiation potential than fertilized embryo-derived embryonic stem cells.展开更多
Focusing on the two themes of"internal control"and"risk management",this paper makes an in-depth analysis of the current situation of risk management and internal control of Moutai Group.It analyze...Focusing on the two themes of"internal control"and"risk management",this paper makes an in-depth analysis of the current situation of risk management and internal control of Moutai Group.It analyzes the current situation of risk management and internal control of Moutai Group.It is found that the risk management of Moutai Group is not perfect,the information exchange is not smooth,and the internal control assessment is not perfect.Finally,it puts forward some corresponding countermeasures,including establishing an effective information communication mechanism,perfecting the risk assessment system,and strengthening the construction of the professional talent team.展开更多
[ Objective] This study aimed to construct nmnmm_ry gland-specific expression vector of bovine tracheal antimicrobial peptide （TAP） gene. [ Method] TAP gene of dairy cattle was amplified from the mammary gland tissu...[ Objective] This study aimed to construct nmnmm_ry gland-specific expression vector of bovine tracheal antimicrobial peptide （TAP） gene. [ Method] TAP gene of dairy cattle was amplified from the mammary gland tissue by RT-PCR using a pair of primers which were designed according to bovine TAP cDNA se- quence （NM_174776） in GenBank, and then cloned into pMD19-T Simple vector for sequencing. The recombinant plasmid was digested using EcoRI and KpnI, the target gene fragment was recovered and inserted into general mammary gland-specific expression vector pBLG-EGFP harboring enhanced green fluorescent protein （ EGFP）, and transfected into bovine mammary epithelial cells （bMEC）, COS-7 cells and lactating rabbit mmmnary gland tissue by lipofectin transfection. The ex- pression of green fluorescent protein in transfected cells was detected under fluorescence microscopy, and the expression of TAP mRNA in rabbit mammary gland tis- sue was detected by semi-quantity RT-PCR. [ Result] The constructed mammary gland-specific expression vector pBLG-EGFP-TAP specifically expressed EGFP in transfected bMECs. In addition, semi-quantitative RT-PCR result showed that the expression level of TAP mRNA in rabbit mammary gland tissue was significantly enhanced after transfeeted with pBLG-EGFP-TAP. [ Conclusion] The mammary gland-specific expression vector pBLG-EGFP-TAP was successfully constructed, which provided important materials for further investigation of expression characteristics of TAP gene and prevention of bovine mastitis by using genetic engineering technology.展开更多
[Objective] This study aimed to prepare dairy cow anti-S100A12 antisem and develop a highly effective and sensitive immunological detection reagent for further investigation of the functions of dairy cow S100A12. [Met...[Objective] This study aimed to prepare dairy cow anti-S100A12 antisem and develop a highly effective and sensitive immunological detection reagent for further investigation of the functions of dairy cow S100A12. [Method] Purified S100A12 protein was respectively emulsified with Freund＇s complete adjuvant and Freund＇s incomplete adjuvant as the antigen for immunizing New Zealand white rabbits to prepare the polyclonal antisera. The titer was detected using agar double diffusion assay and indirect enzyme-linked immunoserbent assay （ELISA） and the specificity was determined with Western Blot. [ Result ] The titer of anti- S100A12 antisera was 1： 8 as determined by agar double diffusion assay and over 1：409 600 by ELISA. Western Blot result showed that the polyclonal antisera could be specifically combined with S100A12 protein. [ Conclusion] The results indicated that anti-S100A12 polyclonal antibody with high fiter and high specificity was successfully obtained, which provided a novel tool for further investigation of the functions of S100A12 gene.展开更多
Background:Non-tuberculous mycobacterial(NTM)infections present in-creasing global health challenges with heterogeneous clinical manifestations and variable immune responses.Despite the growing incidence worldwide,the...Background:Non-tuberculous mycobacterial(NTM)infections present in-creasing global health challenges with heterogeneous clinical manifestations and variable immune responses.Despite the growing incidence worldwide,the molecular mechanisms underlying systemic immune dysfunction in NTM disease remain poorly understood.Methods:We performed comprehensive cross-dataset transcriptomic analysis using two independent RNA-seq da-tasets(GSE97298 and GSE290289)comprising 65 peripheral blood samples from NTM patients and controls.Differential gene expression analysis was conducted using stringent criteria(|log2FC|>1.3,P<0.05),followed by inter-section analysis,protein-protein interaction(PPI)network construction,and functional enrichment analysis.Results:Our analysis identified 10 commonly dysregulated genes across both datasets,forming a highly connected regula-tory network with a network density of 0.267.CD36 emerged as the central hub with the highest degree centrality(0.556)and betweenness centrality(0.722),showing dataset-specific regulation patterns.The network revealed coordinated immune dysfunction characterized by downregulation of T-cell signaling components(CD3E,GZMK)and variable innate immune responses.Functional analysis demonstrated enrichment in pathogen recognition path-ways,lipid metabolism,and inflammatory response regulation.Conclusions:This study provides the first comprehensive cross-dataset analysis of systemic immune networks in NTM disease,identifying CD36 as a central network hub with variable expression patterns.Our findings suggest molecular heterogene-ity in NTM disease and identify potential biomarkers that warrant further val-idation in clinically well-characterized patient cohorts.展开更多
基金supported by the National Natural Science Foundation of China,No. 30900155 and 81070496the Research Foundation of Education Bureau of Shaanxi Province,China,No. 09JK785+1 种基金Foundation of Interdisciplinary for Postgraduates from Northwest University,No. 08YJC22the Key Laboratory Funding of Northwestern University,Shaanxi Province in China
文摘Parthenogenetic embryonic stem cells have pluripotent differentiation potentials, akin to fertilized embryo-derived embryonic stem cells. The aim of this study was to compare the neuronal differentiation potential of parthenogenetic and fertilized embryo-derived embryonic stem cells. Before differentiation, karyotype analysis was performed, with normal karyotypes detected in both parthenogenetic and fertilized embryo-derived embryonic stem cells. Sex chromosomes were identified as XX. Immunocytochemistry and quantitative real-time PCR detected high expression of the pluripotent gene, Oct4, at both the mRNA and protein levels, indicating pluripotent differentiation potential of the two embryonic stem cell subtypes. Embryonic stern cells were induced with retinoic acid to form embryoid bodies, and then dispersed into single cells. Single cells were differentiated in N2 differentiation medium for 9 days. Immunocytochemistry showed parthenogenetic and fertilized embryo-derived embryonic stem cells both express the neuronal cell markers nestin, ~lll-tubulin and myelin basic protein. Quantitative real-time PCR found expression of neuregenesis related genes （Sox-1, Nestin, GABA, Pax6, Zic5 and Pitxl） in both types of embryonic stem cells, and Oct4 expression was significantly decreased. Nestin and Pax6 expression in parthenogenetic embryonic stem cells was significantly higher than that in fertilized embryo-derived embryonic stem cells. Thus, our experimental findings indicate that parthenogenetic embryonic stem cells have stronger neuronal differentiation potential than fertilized embryo-derived embryonic stem cells.
文摘Focusing on the two themes of"internal control"and"risk management",this paper makes an in-depth analysis of the current situation of risk management and internal control of Moutai Group.It analyzes the current situation of risk management and internal control of Moutai Group.It is found that the risk management of Moutai Group is not perfect,the information exchange is not smooth,and the internal control assessment is not perfect.Finally,it puts forward some corresponding countermeasures,including establishing an effective information communication mechanism,perfecting the risk assessment system,and strengthening the construction of the professional talent team.
基金Supported by China Postdoctoral Science Foundation(20090451250)Youth Fund of Sichuan Provincial Department of Education(09zb054)Key Project of International Science and Technology Cooperation(2005DFA30720)
文摘[ Objective] This study aimed to construct nmnmm_ry gland-specific expression vector of bovine tracheal antimicrobial peptide （TAP） gene. [ Method] TAP gene of dairy cattle was amplified from the mammary gland tissue by RT-PCR using a pair of primers which were designed according to bovine TAP cDNA se- quence （NM_174776） in GenBank, and then cloned into pMD19-T Simple vector for sequencing. The recombinant plasmid was digested using EcoRI and KpnI, the target gene fragment was recovered and inserted into general mammary gland-specific expression vector pBLG-EGFP harboring enhanced green fluorescent protein （ EGFP）, and transfected into bovine mammary epithelial cells （bMEC）, COS-7 cells and lactating rabbit mmmnary gland tissue by lipofectin transfection. The ex- pression of green fluorescent protein in transfected cells was detected under fluorescence microscopy, and the expression of TAP mRNA in rabbit mammary gland tis- sue was detected by semi-quantity RT-PCR. [ Result] The constructed mammary gland-specific expression vector pBLG-EGFP-TAP specifically expressed EGFP in transfected bMECs. In addition, semi-quantitative RT-PCR result showed that the expression level of TAP mRNA in rabbit mammary gland tissue was significantly enhanced after transfeeted with pBLG-EGFP-TAP. [ Conclusion] The mammary gland-specific expression vector pBLG-EGFP-TAP was successfully constructed, which provided important materials for further investigation of expression characteristics of TAP gene and prevention of bovine mastitis by using genetic engineering technology.
基金Supported by China Postdoctoral Science Foundation(20090451250)Youth Fund of Sichuan Provincial Department of Education(09ZB054)Program for Changjiang Scholars and Innovative Research Team in University of the Ministry of Education(IRT0848)
文摘[Objective] This study aimed to prepare dairy cow anti-S100A12 antisem and develop a highly effective and sensitive immunological detection reagent for further investigation of the functions of dairy cow S100A12. [Method] Purified S100A12 protein was respectively emulsified with Freund＇s complete adjuvant and Freund＇s incomplete adjuvant as the antigen for immunizing New Zealand white rabbits to prepare the polyclonal antisera. The titer was detected using agar double diffusion assay and indirect enzyme-linked immunoserbent assay （ELISA） and the specificity was determined with Western Blot. [ Result ] The titer of anti- S100A12 antisera was 1： 8 as determined by agar double diffusion assay and over 1：409 600 by ELISA. Western Blot result showed that the polyclonal antisera could be specifically combined with S100A12 protein. [ Conclusion] The results indicated that anti-S100A12 polyclonal antibody with high fiter and high specificity was successfully obtained, which provided a novel tool for further investigation of the functions of S100A12 gene.
文摘Background:Non-tuberculous mycobacterial(NTM)infections present in-creasing global health challenges with heterogeneous clinical manifestations and variable immune responses.Despite the growing incidence worldwide,the molecular mechanisms underlying systemic immune dysfunction in NTM disease remain poorly understood.Methods:We performed comprehensive cross-dataset transcriptomic analysis using two independent RNA-seq da-tasets(GSE97298 and GSE290289)comprising 65 peripheral blood samples from NTM patients and controls.Differential gene expression analysis was conducted using stringent criteria(|log2FC|>1.3,P<0.05),followed by inter-section analysis,protein-protein interaction(PPI)network construction,and functional enrichment analysis.Results:Our analysis identified 10 commonly dysregulated genes across both datasets,forming a highly connected regula-tory network with a network density of 0.267.CD36 emerged as the central hub with the highest degree centrality(0.556)and betweenness centrality(0.722),showing dataset-specific regulation patterns.The network revealed coordinated immune dysfunction characterized by downregulation of T-cell signaling components(CD3E,GZMK)and variable innate immune responses.Functional analysis demonstrated enrichment in pathogen recognition path-ways,lipid metabolism,and inflammatory response regulation.Conclusions:This study provides the first comprehensive cross-dataset analysis of systemic immune networks in NTM disease,identifying CD36 as a central network hub with variable expression patterns.Our findings suggest molecular heterogene-ity in NTM disease and identify potential biomarkers that warrant further val-idation in clinically well-characterized patient cohorts.
