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Objective: To investigate the infection of human embryo fibroblast cell line HF cells by CMV as well as the effects of CMV
on β-actin mRNA and microfilaments. Methods: HF cells shape was observed after the infection of CMV. RT-PCR assay was used
to detect the mRNA expression of CMV immediate early (IE) gene, β-actin and GAPDH genes of HF cells infected by CMV. CMV particles
and cell microfilaments were detected with electron microscope. Results: Shape of HF cell changed after the infection by CMV.
HF cells infected by CMV could express IE mRNA and the expression of β-actin mRNA decreased in a time-and titer-dependent
manner compared with the uninfected HF cells whose expression of GAPDH mRNA did not change much. CMV particles were found
with electron microscope in the cells. Microfilaments were ruptured and shortened after the infection of CMV. Conclusion:
CMV can not only infect human embryo fibroblast cells line HF cells and replicate in the cells, but can also affect the expression
of β-actin mRNA and the microfilaments.
Project (No. 001103058) supported by the Key Program of Science and Technology Bureau of Zhejiang Province, China 相似文献
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肌动蛋白（actin）在维持细胞结构、细胞运动和细胞分裂等过程中发挥着重要的作用.为了克隆鳡（Elopichthys bambusa）β-肌动蛋白（β-actin）全长cDNA,采用反转录PCR（RT-PCR）和cDNA末端快速扩增（rapid amplification of cDNA ends,RACE）技术,从鳡肌肉获得全长为1775bp的β-actin cDNA序列,其中包含1128bp的开放性阅读框,编码375个氨基酸,5′-非翻译（UTR）区为98个核苷酸,3′-UTR区为549个核苷酸.核苷酸与氨基酸的同源性分析发现,鳡β-actin与鲤形目类硬骨鱼的同源性均在98%以上,其中与团头鲂（Megalobrama amblycephala）的同源性最高,分别为98.94%和99.73%,而与其他脊椎类动物如哺乳类、鸟类、两栖类的同源性也分别在85%和97%以上,说明该基因在生物的分子进化过程中具有很高的保守性.采用核苷酸系统发育分析结果显示鳡β-actin与鲤形目类硬骨鱼聚类在一起,且与团头鲂的亲缘关系最近.RT-PCR分析结果显示,β-actin基因在鳡的肝脏、肾、肠、脑、心脏、鳃和肌肉等7个组织均有表达. 相似文献
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目的:研究新城疫病毒(NDV)感染对小鼠肉瘤S180细胞p53蛋白的表达及细胞周期的影响。方法:通过NDV体外感染小鼠肉瘤S180细胞,经倒置显微镜观察细胞形态变化,噻唑蓝(MTT)比色法检测小鼠肉瘤S180细胞的增殖状况;经流式细胞仪分析检测NDV体内感染后荷瘤鼠腹水中S180细胞分裂周期各时相的变化、细胞凋亡情况及细胞表面p53蛋白的表达情况。结果:NDV体内、外感染对小鼠肉瘤S180细胞的杀伤作用明显,NDV体内感染后荷瘤鼠腹水中S180细胞高表达p53蛋白,细胞凋亡率增加,G2/S期细胞减少,增殖指数(PI)降低,与阴性对照组比较有显著性差异(P<0.05)。结论:NDV感染可增强小鼠肉瘤S180细胞p53蛋白的表达,影响其细胞周期,诱导其凋亡且有较强的杀瘤作用。 相似文献
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Yang Li Hang Yan Wu-jun Xue Pu-xun Tian Xiao-ming Ding Xiao-ming Pan Xin-shun Feng Xiao-hui Tian He-li Xiang Jun Hou 《Journal of Zhejiang University. Science. B》2009,10(11):820-828
Objective: To explore the effects of cytomegalovirus (CMV) infection on rejection-related gene expression in the endothelial cells of renal transplantation recipients. Methods: Endothelial cells (ECs) were cultured and stimulated by a variety of factors: A, normal control group; B, inactivated human cytomegalovirus (HCMV) infection group; C, HCMV infection group; D, HCMV supematant infection group; and E, ganciclovir HCMV group. Expression of intercellular adhesion molecule-1 (ICAM-1) and major histocompability complex (MHC) class Ⅰ and class Ⅱ antigens was detected by flow cytometry (FCM) and immuno-histochemistry. Results: We found characteristic CMV-infected ECs in this study. There were no significant differences among groups A, B and D (P>0.05). Although the expression levels of ICAM-1 were not significantly different between groups C and E (P>0.05), the ICAM-1 expression in these two groups was significantly higher than that in group A (P<0.05). ICAM-1 expression was detected in groups C and E, while there was no expression in groups A, B and D. Furthermore, there was no significant difference of ICAM-1 mRNA expression between groups C and E (P>0.05). Human leucocyte antigen (HLA)-ABC expression was detected in all the groups, while HLA-DR expression was only detected in groups C and E. There were no significant dif-ferences of HLA-ABC and HLA-DR expression among groups A, B and D (P>0.05). However, the HLA-ABC and HLA-DR expression levels in groups C and D were higher than those of the remaining groups previously reported (P<0.05). Meanwhile, the HLA-ABC and HLA-DR expression levels in group E were lower than those of group C (P<0.05). Conclusion: CMV could up-regulate the expression levels of ICAM-1 and MHC antigens, which was closely related to allograft rejection. 相似文献
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通过胶原酶、胰蛋白酶双消化法获取蛋鸡松果体细胞,分为对照(培养基)、酒精(溶剂对照)和褪黑素(MT)三组进行原代细胞培养,采用相对定量RT-PCR方法检测褪黑素(MT)对培养的松果体细胞促性腺激素释放激素-Ⅰ(GnRH-Ⅰ)基因表达的影响。结果表明:MTT检测正常培养的松果体细胞增殖速度第9d达高峰,11d缓降。与对照组比较,MT处理1周显著上调其表达丰度(P<0.05);与酒精溶剂组相比,MT处理1周具有上调其表达的趋势(P<0.1);对照组与酒精组之间没有明显的差异(P>0.05)。从而提示:体外培养松果体细胞的GnRH表达受MT的调控。 相似文献
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目的:在康脑液方剂干预下观察皮质区内源性神经细胞干细胞因子(SCF)mRNA在大鼠脑缺血再灌注损伤后不同时间的表达情况。方法:成年SD大鼠,以线栓法建立大脑中动脉缺血再灌注模型,随机分为模型组、药物干预组和假手术组。原位杂交技术检测脑缺血1.5h再灌注1～14d后,脑皮质区SCFmRNA表达情况。结果:脑缺血再灌注后,药物干预组、模型组SCFmRNA的表达在皮质区均明显高于假手术组,于第7天达高峰,第14天下降。结论:药物干预后脑缺血再灌注脑皮质区SCFmRNA表达在不同时间点与模型组具备相同的表达规律,两组SCFmRNA的表达无显著性差异。 相似文献
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Lyu Sunjian Yuan Xuemei Liu Li Zhang Haiqi Yu Zhe Hang Xiaoying Shi Weida Wu Yinglei 《Journal of Zhejiang University. Science. B》2021,22(4):295-304
Trionyx sinensis Hemorrhagic Syndrome Virus(TSHSV) is an arterivirus newly discovered in Chinese softshell turtles. Little is known about the effect of antibodies against the virus or the distribution of the virus in different organs of infected turtles. In this study, a partial protein of TSHSV-HP4 was produced using a prokaryotic expression system, and its polyclonal antibody was generated. The polyclonal antibody was confirmed by western blot and dot enzyme-linked immunosorbent assay(dot-ELISA). The distribution of TSHSV in different organs of T. sinensis was examined by immunohistochemistry(IHC) and the expression of immune-related genes was analyzed using quantitative real-time polymerase chain reaction(qRT-PCR). The results indicated that the recombinant TSHSV-HP4 protein was successfully expressed, and the generated polyclonal antibody showed specific binding to viral particles in the lung tissues of infected turtles. The IHC assay indicated that the virus was highly localized in various cells, including intestinal lymphocytes,enterocytes, kidney epithelial cells, spleen cells, lung macrophages, and cardiomyocytes. The qRT-PCR analysis revealed that TSHSV was detected in all organs tested, including the lungs, liver, kidneys, spleen, and heart. The numbers of viral mRNA copies in lung and heart tissues were significantly higher in the virus-antibody group than in the virus group. The interferonstimulated genes(ISGs), myxovirus resistance protein 2(MX2) and radical S-adenosyl methionine domain containing 2(RSAD2) were highly upregulated in all groups of infected turtles. Antibody-dependent enhancement(ADE) seemed to occur after stimulation by the polyclonal antibody, because significantly greater expression of the two genes was detected in the virus-antibody group than in the virus group. Overall, these results are important in understanding the cell localization of TSHSV and the immune response of infected turtles. 相似文献
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INTRODUCTION Recently discovered as an endocrine organ, adi-pose tissue can secrete many cytokines such as Re-sistin, adiponectin and free fatty acids, and receives much attention in the study of insulin resistance and type 2 diabetes mellitus (T2DM). As a peptide hor-mone secreted by adipocyte, Resistin is considered to be the linkage between obesity and insulin resistance (Steppan et al., 2001). Most studies on Resistin in-vestigate adipose tissue. In this experiment we de-tected t… 相似文献
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采用实时荧光定量PCR技术,以β-actin为内参,检测了白羽AA肉鸡在4周龄时其胸肌、腿肌、脂肪、心、肝、脾、肺、肾、腺胃、肌胃、肠、脑共12个组织和在1日龄及1、2、3、4、5、6、7周龄时腿肌、肝脏、脂肪中类胰岛素生长因子-I（IGF-I）的表达量变化。结果显示,除肾外其余组织均表达一定量的IGF-I mRNA表达,且在肝脏中表达量显著高于其他组织;腿肌6周时IGF-I mRNA表达量达到最大值,其余各时期也都高于1日龄时的表达;肝脏中表达量最高时期是3周,然后又呈下降趋势;脂肪中在3周时表达量最高,然后其表达量随年龄增长而逐渐下降。本实验结果揭示了生长早期肉鸡IGF-I基因mRNA的时空表达特征,为进一步研究生长轴相关基因对肉鸡早期生长发育的调控作用奠定了基础。 相似文献
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Sun Jie Huang He Zhu Yuan-yuan Lan Jian-ping Li Jing-yuan Lai Xiao-yu Yu Jian 《Journal of Zhejiang University. Science. B》2005,6(12):1141-1147
Objective: Detecting the expression and mutation of human telomeric repeat binding factor (hTRF1) in 10 malignant hematopoietic cell line cells on the base of determining its genomic structure and its four pseudogenes
to clarify if hTRF1 mutation is one of the factors of the activation of telomerase. Methods: hTRF1cDNA sequences were obtained from GenBank, its genome structure and pseudogenes were forecasted by BLAST and other biology
information programs and then testified by sequencing. Real-time RT-PCR was used to detect the expression of hTRF1mRNA in 10 cell line cells, including myelogenous leukemia cell lines K562, HL-60, U-937, NB4, THP-1, HEL and Dami; lymphoblastic
leukemia cell lines 6T-CEM, Jurkat and Raji. Telomerase activities of cells were detected by using telomeric repeat amplification
(TRAP)-ELISA protocol. PCR and sequencing were used to detect mutation of each exon of hTRF1 in 10 cell line cells. Results: hTRF1 gene, mapped to 8q13, was divided into 10 exons and spans 38.6 kb. Four processed pseudogenes of hTRF1 located on chromosome 13, 18, 21 and X respectively, was named as ψhTRF1-13, ψhTRF1-18, ψhTRF1-21 and ψhTRF1-X respectively. All cell line cells showed positive telomerase activity. The expression of hTRF1 was significantly lower in malignant hematopoietic cell lines cells (0.0338, 0.0108:_0.0749) than in normal mononuclear cells
(0.0493, 0.0369:_0.128) (P=0.004). But no significant mutation was found in all exons of hTRF1 in 10 cell line cells. Four variants were found in part of intron 1, 2 and 8 of hTRF1. Their infection on gene function is unknown and needs further studies. Conclusion: hTRF1 mutation is probably not one of the main factors for telomerase activation in malignant hematopoietic disease.
Project supported by the National Basic Research Program (973) of China (No. 2002CB713700) and the National Natural Science
Foundation of China (No. 39870339) 相似文献
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Lang QL Zhou XC Zhang XL Drabek R Zuo ZX Ren YL Li TB Chen JS Gao XL 《Journal of Zhejiang University. Science. B》2011,12(2):116-125
A large number of plant microRNAs (miRNAs) are now documented in the miRBase, among which only 30 are for Solanum lycopersicum (tomato). Clearly, there is a far-reaching need to identify and profile the expression of miRNAs in this important crop under
various physiological and pathological conditions. In this study, we used an in situ synthesized custom microarray of plant
miRNAs to examine the expression and temporal presence of miRNAs in the leaves of tomato plants infected with Cucumber mosaic virus (CMV). Following computational sequence homology search and hairpin structure prediction, we identified three novel tomato
miRNA precursor genes. Our results also show that, in accordance with the phenotype of the developing leaves, the tomato miRNAs
are differentially expressed at different stages of plant development and that CMV infection can induce or suppress the expression
of miRNAs as well as up-regulate some star miRNAs (miRNA*s) which are normally present at much lower levels. The results indicate
that developmental anomalies elicited by virus infection may be caused by more complex biological processes. 相似文献
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为了解无孢子生殖胚囊助细胞的特点,用透射电子显微镜对无孢子生殖植物非洲狼尾草无融合生殖胚囊的助细胞进行了研究.结果表明,助细胞壁产生无数突起,伸入细胞质形成巨形丝状器,呈圆锥形,细胞质深入丝状器内部,这一结构对引入花粉管有利;细胞含在量线粒体、高尔基体、核糖体等细胞器,以丝状器周围最为丰富,显示出旺盛的代谢活动;具有丰富的贮藏物质,先是以淀粉形式积累,然后淀粉转化为脂类物质,以脂滴形式存在,细胞解体后供卵和中央细胞利用.无孢子生殖胚囊肋细胞的结构特点与有性生殖胚囊助细胞一致,其存在为花粉管进入胚囊提供通道,并为胚囊其它成员发育提供营养. 相似文献
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为了了解能恢复籼粳杂种一代育性的广亲和基因在植物体内作用的部位和时间,本文应用扫描电镜对小花芽分化到小花形成的全过程,并通过整体染色及透明法,显微切片,借相差显微镜及微分干涉显微镜,对籼、粳稻及3个籼粳杂种F_1代的材料进行了观察和研究。结果表明:在杂种的大孢子发生过程中,可观察到多种形式的细胞解体过程,可导致无胚囊子房的形成,最后造成小花的敗育。因而认为,广亲和基因的作用很可能与杂种的胚囊发生,特别是大孢子发生中的异常过程的纠正有关。 相似文献
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Fu Y Wang SQ Liu YP Wang GP Wang JT Gong SS 《Journal of Zhejiang University. Science. B》2008,9(4):299-305
Objective: To evaluate the transduction efficiency of a recombinant adenovirus carrying the gene for green fluorescent protein (Ad-GFP) into the primary cultures of fetal neural stem cells (NSCs) by the expression of GFP. Methods: The Ad-GFP was constructed by homologous recombination in bacteria with the AdEasy system; NSCs were isolated from rat fetal hippocampus and cultured as neurosphere suspensions. After infection with the recombinant Ad-GFP, NSCs were examined with a fluorescent microscopy and a flow cytometry for their expression of GFP. Results: After the viral infection, flow cytometry analysis revealed that the percentage of GFP-positive cells was as high as 97.05%. The infected NSCs sustained the GFP expression for above 4 weeks. After differentiated into astrocytes or neurons, they continued to express GFP efficiently. Conclusion: We have successfully constructed a viral vector Ad-GFP that can efficiently infect the primary NSCs. The reporter gene was showed fully and sustained expression in the infected cells as well as their differentiated progenies. 相似文献
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目的 :研究新城疫病毒在体外抗胃癌细胞活性及其与细胞凋亡的关系。方法 :应用倒置显微镜观察细胞形态、MTT法测NDV在体外对BGC - 82 3的抑制和杀伤作用 ,同时用流式细胞术检测胃癌细胞凋亡情况及细胞分裂周期各时象的变化。结果 :NDV在体外可使BGC - 82 3胃癌细胞形成明显的细胞病变效应、细胞生长抑制及细胞凋亡 ,且细胞凋亡率与感染时间呈正相关。G2、S期细胞减少 ,增殖指数 (PI)降低 ,与阴性对照组比较有显著性差异 (P<0 .0 1)。结论 :NDV具有显著的抗BGC - 82 3胃癌细胞活性。 相似文献
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通过石蜡切片,汞-溴酚蓝染色,在显微镜下系统观察了川白芷胚胎发育时期蛋白质的动态变化.结果表明:在受精后1-16d,蛋白质积累逐渐增多,且胚细胞中的蛋白质含量的增加明显快于珠被;合点端细胞中蛋白质很明显,珠孔端蛋白质少于合点端;第17d后,胚和珠被的蛋白质含量逐渐下降. 相似文献
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Proapoptotic and pronecrosis effect of different truncated hepatitis C virus core proteins 总被引：2，自引：0，他引：2
Yan XB Chen Z Luo DH Xu XY Wu W Zhou LF 《Journal of Zhejiang University. Science. B》2005,6(4):295-300
Objective: To study the roles of different truncated hepatitis C virus (HCV) core proteins (CORE) in the pathogenesis of HCV persistent infection and hepatocellular carcinoma (HCC) and to assess intracellular localization in transiently transfected cells. Methods: Seven truncated CORE-GFP (green fluorescent protein) fusion protein expression plasmids were constructed,which contained HCV CORE sequences derived from tumor tissues (BT) and non-tumor tissues (BNT) from one patient infected with HCV. Amino acid (aa) lengths were BT: 1-172 aa, 1-126 aa, 1-58 aa, 59-126 aa, 127-172 aa; BNT: 1-172 aa and C191:1-172 aa respectively. Subcellular localization of CORE-GFP was analyzed by con-focal laser scanning microscope. Apoptosis and necrosis were quantified by flow cytometry. Results: Different truncated CORE-GFP localized mainly in the cytoplasm, but nuclear staining was also observed. HCV CORE could induce apoptosis and necrosis, and different truncated COREs could induce cell apoptosis and necrosis at different levels. Among the same length 1-172 aa of BT, BNT and C191, the cell apoptosis and necrosis percentage of BT is highest, and C191 is the lowest (BT＞BNT＞C191). To the different fragment COREs of BT,N-terminal of CORE induced apoptosis and necrosis higher, compared with that of C-terminal (1-172 aa＞1-126 aa＞1-58aa＞ 127-172 aa＞59-126 aa). Conclusion: These results suggest HCV CORE could induce apoptosis and necrosis of cells, which might play an important role in the pathogenesis of HCV persistent infection and HCC and the different CORE domains of different HCV quasi-species might have some difference in their pathogenesis. 相似文献