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To construct a hepatic stellate cells (HSCs) subtracted cDNA library to find differentially expressed genes in normal mice and mice infected with Schistosomajaponicum (S. japonicum). Suppression subtractive hybridization (SSH) was used. The cDNA fragments of normal mouse were compared to those of schistosoma-infected mice to find differentially expressed genes. Then differentially expressed cDNA fragments were directly inserted into T/A cloning vector to set up the subtractive library. Amplification of the library was carried out with transformation of DH5α. The amplified library contained more than 400 positive bacterial clones, which were then hybridized with forward and backward subtracted probes for differential screening, One hundred positive bacterial clones were randomly selected for sequencing and BLAST analysis, Finally, virtual Northern Blot confirmed such differential expression. The subtracted cDNA library of differentially expressed genes of HSCs was constructed successfully, the library is efficient and lays foundation for screening and cloning new and specific genes of schistosomiasis.  相似文献   

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为研究海洋防污损涂料添加剂Irogarol-1051的降解产物M2的基因毒性,应用微阵列技术,选取Affymetrix公司鼠基因组230 2.0基因芯片检测30μmol/L M2暴露下的鼠肝癌细胞基因表达变化.实验结果显示,96 h的M2暴露导致了38个基因在全部4组可能的对照/暴露中均发生显著变化(p0.002 5),其中只有Accn5基因研究较为透彻,该基因表达的抑制可能影响上皮钠通道的功能.此外,分别有10和82个功能注释基因在至少一组对照/暴露组中上调和下调.M2诱导的基因主要和细胞核(细胞成分)相关.M2抑制的基因则主要影响生物过程中的G蛋白偶联信号通路功能和细胞成分中的细胞膜内整合功能.  相似文献   

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Objective

The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipidassociated membrane proteins (LAMPs) in the host innate immune system, specifically their effect on Toll-like receptors (TLRs).

Methods

LAMPs were derived from U. urealyticum strains, and human amniotic epithelial cells (HAECs) were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay (ELISA) and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry.

Results

LAMPs induced HAECs to produce inflammatory cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor (anti-hTLR2-IgA).

Conclusions

LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.
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Objective:To find new protein biomarkers for the detection and evaluation of liver injury and to analyze the relationship between such proteins and disease progression in concanavalin A (Con A)-induced hepatitis.Methods:Twenty-five mice were randomly divided into five groups:an untreated group,a control group injected with phosphate buffered saline (PBS),and groups with Con A-induced hepatitis evaluated at 1,3 and 6 h.Two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) were used to identif...  相似文献   

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Objective: To find new protein biomarkers for the detection and evaluation of liver injury and to analyze the relationship between such proteins and disease progression in concanavalin A (Con A)-induced hepatitis. Methods: Twenty-five mice were randomly divided into five groups: an untreated group, a control group injected with phosphate buffered saline (PBS), and groups with Con A-induced hepatitis evaluated at 1, 3 and 6 h. Two-dimensional gel elec-trophoresis (2-DE) and mass spectrometry (MS) were used to identify differences in protein expression among groups. Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to verify the results. Results: In mice with Con A-induced hepatitis, expression levels of four proteins were increased: RIKEN, fructose bisphosphatase 1 (fbp1), ketohexokinase (khk), and Chain A of class pi glutathione S-transferase. Changes in fbp1 and khk were confirmed by qRT-PCR. Conclusion: Levels of two proteins, fbp1 and khk, are cleady up-regulated in mice with Con A-induced hepatitis.  相似文献   

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Epstein-Barr virus(EBV),a human gammaherpesvirus carried by more than 90% of the world’s population,is associated with malignant tumors such as Burkitt’s lymphoma(BL),Hodgkin lymphoma,post-transplant lymphoma,extra-nodal natural killer/T cell lymphoma,and nasopharyngeal and gastric carcinomas in immune-compromised patients.In the process of infection,EBV faces challenges:the host cell environment is harsh,and the survival and apoptosis of host cells are precisely regulated.Only when host cells receive sufficient survival signals may they immortalize.To establish efficiently a lytic or long-term latent infection,EBV must escape the host cell immunologic mechanism and resist host cell apoptosis by interfering with multiple signaling pathways.This review details the apoptotic pathway disrupted by EBV in EBV-infected cells and describes the interactions of EBV gene products with host cellular factors as well as the function of these factors,which decide the fate of the host cell.The relationships between other EBV-encoded genes and proteins of the B-cell leukemia/lymphoma(Bcl) family are unknown.Still,EBV seems to contribute to establishing its own latency and the formation of tumors by modifying events that impact cell survival and proliferation as well as the immune response of the infected host.We discuss potential therapeutic drugs to provide a foundation for further studies of tumor pathogenesis aimed at exploiting novel therapeutic strategies for EBV-associated diseases.  相似文献   

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Two novel sugar-conjugated 5-fluorocytosine (5-FC) antineoplastic compounds were designed and synthesized to improve the selective drug uptake by targeting the tumor-specific glucose transporter (GLUT).The antitumor activity of these compounds was evaluated in four different human cancer cell lines:A549 (human lung cancer cell line),HT29 (human colorectal cancer cell line),H460 (human lung cancer cell line),and PC3 (human prostate cancer cell line).The sugar conjugates exhibited cytotoxicity similar to or higher than 5-FC and 1-hexylcarbamoyl-5-FC in A549,HT29,H460,and PC3.Furthermore,GLUT-mediated transport of the glycoconjugate was investigated with GLUT inhibitor-mediated cytotoxicity analysis in a GLUT-overexpressing HT29 cell line.The cell-killing potency of 5-FC glycoconjugate was found to depend significantly on the GLUT inhibitor,and the cellular uptake of molecules was regulated by GLUT-mediated transport.All the results demonstrate the potential advantages of glycoconjugation for Warburg effect-targeted drug design.  相似文献   

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目的:基于网络药理学方法探讨威灵仙抗结直肠癌可能的分子机制。方法:通过TCMSP数据库挖掘获取威灵仙活性成分,限定口服利用度(OB)和类药性(DL)数值对威灵仙活性成分进行筛选。利用TCMSP搜索威灵仙相关靶基因,GeneCards数据库收集与结直肠癌相关的基因,采用Cytoscape Version 3.7.1软件,构建威灵仙活性成分-抗结直肠癌靶点网络。使用String数据库构建蛋白质相互作用网络。使用Bioconductor数据库对威灵仙抗结直肠癌作用靶点进行基因本体(GO)富集分析与KEGG通路分析,以探究威灵仙抗结直肠癌的作用机制。结果:收集到57种化合物,经筛选和分析获得威灵仙活性成分7个,涉及抗结直肠癌作用靶点30个,关键靶点包括CASP3、JUN、ADRA1B、MAOA等。GO功能富集分析得到GO条目55个,KEGG通路富集筛选得到61条信号通路,包括凋亡通路、p53信号通路、结直肠癌信号通路等。结论:威灵仙治疗结直肠癌具有多通路、多靶点作用的特点,其具体的调控机制及关键的靶标尚待通过进一步的实验研究加以探索及验证。  相似文献   

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INTRODUCTION 1,25-dihydroxyvitamin D3[1,25(OH)2D3], the biologically active metabolite of vitamin D3, is a secosteroid hormone that not only regulates bone and calcium/phosphate metabolism but also regu-lates a number of other biological activities, in-cluding modulation of the immune response via specific receptors expressed in antigen presenting cells (APC) and activated T cells. Recently, in-creasing evidence showed that the modulatory role of 1,25(OH)2D3 on T cell phenotype and …  相似文献   

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目的:寻找来源于Alport综合征患者与正常对照者的多能干细胞差异性表达的microR NA,并对差异性表达的microR NA靶基因进行预测。创新点:本研究不同于一般的试验标本,试验标本是来源于尿肾脏管细胞诱导而成的多能干细胞。基于Alport综合征是遗传疾病,我们对一遗传家系进行了系统的分析。寻找特异性的差异性表达microR NA及其靶基因,从基因水平对Alport综合征进行研究。方法:在前期工作中,成功地从实验者与对照者的尿肾脏管细胞诱导成多能干细胞。运用高通量测序技术分析并发现实验者与对照者之间差异性表达的microR NA。对差异性表达的microR NA靶基因进行预测,并进行靶基因聚集分析,研究靶基因主要参与的生物学过程。同时进行靶基因信号传导通路的分析,研究靶基因主要参与的细胞信号传导通路。结论:在实验组与对照组之间,发现了30个具有显著差异性表达的microR NAs,包括19个上调表达与11个下调表达。差异性表达的microR NA的靶基因主要聚集在细胞膜和细胞代谢过程;靶基因主要参与嘌呤代谢通路与丝裂原活化蛋白激酶(MAPK)通路。  相似文献   

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The aim of this study was to research the changes in cytotoxicity and antibacterial properties after silver nanoparticles (AgNPs) were incorporated into the surface coating of dental alloys. AgNPs were attached to cobalt chromium alloys and pure titanium using a hydrothermal method, according to the reaction: AgNO3+NaBH4→ Ag+1/2H2+1/2B2H6+NaNO3. A 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to evaluate the cytotoxicity of the alloys when in contact with osteogenic precursor cells (MC3T3-E1) from mice and mesenchymal stem cells (BMSC) from rats. The antibacterial properties of dental alloys incorporating three different concentrations (10, 4, and 2 μmol/L) of AgNPs were tested on Staphylococcus aureus (SA) and Streptococcus mutans (MS). High cytotoxicity values were observed for all dental alloys that contained 0% of AgNPs (the control groups). The incorporation of AgNPs reduced cytotoxicity values. No significant difference was observed for antibacterial performance when comparing dental alloys containing AgNPs to the respective control groups. The results demonstrated that the cobalt chromium alloys and pure titanium all had cytotoxicity to MC3T3-E1 and BMSC and that the incorporation of AgNPs could reduce this cytotoxicity. The concentrations of AgNPs adopted in this study were found to have no antibacterial action against SA or MS.  相似文献   

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Objective: To determine whether spinal cord decompression plays a role in neural cell apoptosis after spinal cord injury. Study design: We used an animal model of compressive spinal cord injury with incomplete paraparesis to evaluate neural cell apoptosis after decompression. Apoptosis and cellular damage were assessed by staining with terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate nick-end labelling (TUNEL) and immunostaining for caspase-3, Bcl-2 and Bax. Methods: Experiments were conducted in male Spragne-Dawley rats (n=78) weighing 300-400 g. The spinal cord was compressed posteriorly at T10 level using a custom-made screw for 6 h, 24 h or continuously, followed by decompression by removal of the screw. The rats were sacrificed on Day 1 or 3 or in Week 1 or 4 post-decompression. The spinal cord was removed en bloc and examined at lesion site, rostral site and caudal site (7.5 mm away from the lesion). Results: The numbers of TUNEL-positive cells were significantly lower at the site of decompression on Day l, and also at the rostral and caudal sites between Day 3 and Week 4 post-decompression, compared with the persistently compressed group. The numbers of cells between Day 1 and Week 4 were immunoreactive to caspase-3 and B-cell lymphoma-2 (Bcl-2)-associated X-protein (Bax), but not to Bcl-2, correlated with those of TUNEL-positive cells. Conclusion: Our results suggest that decompression reduces neural cell apoptosis following spinal cord injury.  相似文献   

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