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991.
研究鉴定激活hfgl2凝血酶原酶基因的SARS冠状病毒结构蛋白.从SARS尸检肺组织中抽提RNA后制备cDNA,分别扩增SARS-CoV的N、S2和M全长基因序列,再分别克隆到真核表达载体pcDNA3.1(+)上.应用免疫组织化学分析鉴定pcDNA3.1-N、pcDNA3.1-M和pcDNA3.1-S2的表达.构建人纤维介素(hfgl2)启动子荧光素酶报告基因质粒,并将SARS冠状病毒结构蛋白表达质粒分别与其共转染以明确激活hfgl2基因转录的SARS冠状病毒结构蛋白.将目的片段克隆至pcDNA3.1(+),经酶切鉴定和测序鉴定无误;免疫组织化学染色可见明显的CHO细胞胞浆棕染.与hfgl2启动子共转染实验阐明SARS冠状病毒膜(M)蛋白和刺突糖(S2)蛋白对hfgl2基因的激活与对照组无显著差异,而SARS冠状病毒核心(N)蛋白可激活hfgl2启动子,使其转染活性提高4.6倍.SARS冠状病毒N蛋白可增强hfgl2基因的转录活性. 相似文献
992.
Tianyu Liu Zixuan Guo Xueli Song Li Liu Wenxiao Dong Sinan Wang Mengque Xu Cheng Yang Bangmao Wang Hailong Cao 《Journal of cellular and molecular medicine》2020,24(4):2648-2662
High‐fat diet (HFD) is a well‐known risk factor for gut microbiota dysbiosis and colorectal cancer (CRC). However, evidence relating HFD, gut microbiota and carcinogenesis is limited. Our study aimed to demonstrate that HFD‐induced gut dysbiosis promoted intestinal adenoma‐adenocarcinoma sequence. In clinical study, we found that HFD increased the incidence of advanced colorectal neoplasia (AN). The expression of monocyte chemoattractant protein 1 (MCP‐1), CC chemokine receptor 2 (CCR2) and CD163 in CRC patients with HFD was significantly higher than that in CRC patients with normal diet. When it comes to the Apcmin/+ mice, HFD consumption could induce gut dysbiosis and promote intestinal carcinogenesis, accompanying with activation of MCP‐1/CCR2 axis that recruited and polarized M2 tumour‐associated macrophages. Interestingly, transfer of faecal microbiota from HFD‐fed mice to another batch of Apcmin/+ mice in the absence of HFD could also enhance carcinogenesis without significant body weight gain and induced MCP‐1/CCR2 axis activation. HFD‐induced dysbiosis could also be transmitted. Meanwhile, antibiotics cocktail treatment was sufficient to inhibit HFD‐induced carcinogenesis, indicating the vital role of dysbiosis in cancer development. Conclusively, these data indicated that HFD‐induced dysbiosis accelerated intestinal adenoma‐adenocarcinoma sequence through activation of MCP‐1/CCR2 axis, which would provide new insight into better understanding of the mechanisms and prevention for HFD‐related CRC. 相似文献
993.
An Evaluation of the Impact of PD‐1 Pathway Blockade on Reproductive Safety of Therapeutic PD‐1 Inhibitors 下载免费PDF全文
Frederique M. Poulet Jayanthi J. Wolf Danuta J. Herzyk Joseph J. DeGeorge 《Birth defects research. Part B, Developmental and reproductive toxicology》2016,107(2):108-119
This report discusses the principles of reproductive toxicity risk assessment for biopharmaceuticals blocking the PD‐1/programmed cell death ligand 1 (PD‐L1) pathway, which have been developed for the treatment of patients with advanced malignancies. The PD‐1/PD‐L1 pathway is a T‐cell co‐inhibitory pathway that normally maintains immune tolerance to self. Its role in pregnancy is to maintain immune tolerance to the fetal allograft. In cancer patients, this signaling pathway is hijacked by some neoplasms to avoid immune destruction. PD‐1/PD‐L1‐blocking agents enhance functional activity of the target lymphocytes to eventually cause immune rejection of the tumor. A therapeutic blockade of PD‐1/PD‐L1 pathway that occurs at full target engagement provides a unique challenge to address the risk to pregnancy because disruption of the same pathway may also reduce or abrogate maternal immune tolerance to the fetal alloantigens inherited through the father. Typically, nonclinical reproductive and developmental toxicity (DART) studies in animals (rats and rabbits) with clinical drug candidates are conducted to identify potential risk in humans and to determine exposure margin for the effects on reproduction as part of the risk assessment. However, for biopharmaceuticals for which the desired mechanism of action cannot be separated from potential deleterious effects to the fetus and when the only relevant toxicology species is nonhuman primate (NHP), the risk to reproduction can be predicted by a mechanism‐based assessment using data generated from murine surrogate models as supportive information without conducting DART in NHPs. Such an approach has been used in the evaluation of pregnancy risk of anti‐PD‐1 agent, pembrolizumab, and has been demonstrated as an important alternative to performing DART studies in NHPs 相似文献
994.
Douglas?R.?Boettner Christopher?Huston William?A.?PetriEmail author 《Journal of biosciences》2002,27(6):553-557
Entamoeba histolytica is an enteric parasite that can kill host cells via a contact-dependent mechanism. This killing involves the amoebic surface
protein referred to as the Gal/GalNAc lectin. The Gal/GalNAc lectin binds galactose and N-acetylgalactosamine allowing the
adherence of amoebas to host cells. Involvement of the lectin in the pathogenesis ofE. histolytica infection will be reviewed in this paper. The lectin has been shown to have very specific and substantial effects on adherence,
cytotoxicity, and encystation. There is also possible involvement of the lectin in phagocytosis and caspase activation in
host cells. 相似文献
995.
杜氏盐藻rbcS启动子的克隆和功能分析 总被引:2,自引:0,他引:2
为提高转基因盐藻的表达效率,利用基因组步行方法和巢式PCR,从盐藻中克隆了1,5-二磷酸核酮糖羧化酶/加氧酶(Rubisco)的小亚基基因rbcS 的5'上游调控序列,并对其进行序列分析和转基因功能分析。采用Dra I、EcoR V、Pvu II和Stu I四种平端限制内切酶分别酶切盐藻基因组DNA,并与接头连接,构建基因组步行文库GWL 1、GWL 2、GWL 3和GWL 4;设计特异引物从这四种文库中扩增rbcS基因的5'上游调控序列。在GWL 1、GWL 4中分别扩增出约1.2 kb的片段。对该序列的分析表明,它的3'端与已知盐藻rbcS cDNA 的5'端序列完全一致,说明是该基因的5'端上游区,并且包含多个与转录调控有关的保守序列(如TATA-box、CAAT-box),富含GT的重复序列。此序列EcoR I下游的片段与除草剂抗性基因bar相融合,构建表达载体,电击法转化盐藻。通过对转化藻株的抗性筛选以及PCR和Southern blot检测,表明该区域能驱动外源基因bar在转基因盐藻中的表达,推断是盐藻rbcS基因的启动子调控区。 相似文献
996.
为探究自噬抑制剂6-氨基-3-甲基腺嘌呤(3-methyladenine,3-MA)对损伤细胞氧化应激水平的影响,将3-MA作用于H2O2诱导的PC12细胞损伤模型,以自噬增强剂雷帕霉素(rapamycin,Rap)作为对照,探讨自噬与氧化应激的关系。测定线粒体的膜电位和细胞内的活性氧(reactive oxygen species, ROS)与丙二醛(malondialdehyde, MDA)含量,以及超氧化物歧化酶(superoxide dismutase,SOD)和过氧化氢酶(catalase,CAT)活性,评价损伤细胞的氧化应激状态。单丹(磺)酰戊二胺(monodansylcadaverine,MDC)染色,观察损伤细胞的自噬情况。蛋白质印迹分析损伤细胞中的自噬相关蛋白质LC3-II/LC3-I比值变化。实验结果显示:与正常组相比,H2O2损伤细胞的ROS水平上升到正常组的141%,MDA含量增加(P<0.001);CAT与SOD酶活力显著降低(P<0.001),差异均有统计学意义,证明损伤细胞氧化应激水平增加;MDC染色结果表明,H2O2组自噬明显增加。Western印迹结果表明,LC3-II/LC3-I值显著升高(P<0.05);与损伤组相比,3-MA组MDC染色结果表明,自噬水平降低。Western印迹结果表明,LC3-II/LC3-I值下降;细胞内ROS水平升高,增加到正常组的208%。MDA含量增加(P<0.001),CAT、SOD酶活力降低(P<0.001)。综上结果表明,自噬抑制剂可增加H2O2诱导的PC12细胞损伤模型的氧化应激水平,增加细胞凋亡。 相似文献
997.
Identification and expansion of pancreatic stem/progenitor cells 总被引:4,自引:0,他引:4
Pancreatic islet transplantation represents an attractive approach for the treatment of diabetes. However, the limited availability of donor islets has largely hampered this approach. In this respect, the use of alternative sources of islets such as the ex vivo expansion and differentiation of functional endocrine cells for treating diabetes has become the major focus of diabetes research. Adult pancreatic stem cells /progenitor cells have yet to be recognized because limited markers exist for their identification. While the pancreas has the capacity to regenerate under certain circumstances, questions where adult pancreatic stem/progenitor cells are localized, how they are regulated, and even if the pancreas harbors a stem cell population need to be resolved. In this article, we review the recent achievements both in the identification as well as in the expansion of pancreatic stem/progenitor cells. 相似文献
998.
Deep brain stimulation 总被引:9,自引:0,他引:9
During the last decade deep brain stimulation (DBS) has become a routine method for the treatment of advanced Parkinsons disease (PD), leading to striking improvements in motor function and quality of life of PD patients. It is associated with minimal morbidity. The rationale of targeting specific structures within basal ganglia such as the subthalamic nucleus (STN) or the internal segment of the globus pallidus (GPi) is strongly supported by the current knowledge of the basal ganglia pathophysiology, which is derived from extensive experimental work and which provides the theoretical basis for surgical therapy in PD. In particular, the STN has advanced to the worldwide most used target for DBS in the treatment of PD, due to the marked improvement of all cardinal symptoms of the disease. Moreover on-period dyskinesias are reduced in parallel with a marked reduction of the equivalent daily levodopa dose following STN–DBS. The success of the therapy largely depends on the selection of the appropriate candidate patients and on the precise implantation of the stimulation electrode, which necessitates careful imaging-based pre-targeting and extensive electrophysiological exploration of the target area. Despite the clinical success of the therapy, the fundamental mechanisms of high-frequency stimulation are still not fully elucidated. There is a large amount of evidence from experimental and clinical data that stimulation frequency represents a key factor with respect to clinical effect of DBS. Interestingly, high-frequency stimulation mimics the functional effects of ablation in various brain structures. The main hypotheses for the mechanism of high-frequency stimulation are: (1) depolarization blocking of neuronal transmission through inactivation of voltage dependent ion-channels, (2) jamming of information by imposing an efferent stimulation-driven high-frequency pattern, (3) synaptic inhibition by stimulation of inhibitory afferents to the target nucleus, (4) synaptic failure by stimulation-induced neurotransmitter depletion. As the hyperactivity of the STN is considered a functional hallmark of PD and as there is experimental evidence for STN-mediated glutamatergic excitotoxicity on neurons of the substantia nigra pars compacta (SNc), STN–DBS might reduce glutamatergic drive, leading to neuroprotection. Further studies will be needed to elucidate if STN–DBS indeed provides a slow-down of disease progression. 相似文献
999.
WuQiang F Yanase T Wei L Oba K Nomura M Okabe T Goto K Nawata H 《Biochemical and biophysical research communications》2003,311(4):987-994
Ad4BP/SF-1 plays key roles at all levels of the hypothalamic-pituitary-steroidogenic organ axis and its functional disruption causes endocrine disorders of these organs. However, only three human subjects with Ad4BP/SF-1 mutations have been reported to date, suggesting limited clinical significance as a cause of inborn adrenal or sexual abnormalities. We report the first functional characterization of a new variation found in the hinge region of human Ad4BP/SF-1, G146A. Resulting from a single nucleotide shift (GGG-->GCG), G146A bears slightly diminished transactivation activity evidenced by both adrenal specific cyp11A promoter and ovary specific cyp19 promoter II. The variation does not affect protein expression or stability, exhibiting no dominant negative effect. G146A has a normal interaction pattern with standard co-regulators and subnuclear distribution pattern, and can be considered as a nonsynonymous single nucleotide polymorphism, since it occurs in normals and patients with adrenal diseases. In normal Japanese the allele C frequency is 8%, while in a preliminary population of patients with adrenal diseases it is elevated to 30%; suggesting the G146A variation might be of clinical importance. 相似文献
1000.
Specific molecular marker of the genes controlling linolenic acid content in rapeseed 总被引:4,自引:0,他引:4
C. Jourdren P. Barret D. Brunel R. Delourme M. Renard 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,93(4):512-518
In rapeseed, which is an agronomically important oilseed, variation in the linolenic acid content of the oil has been obtained through chemical mutagenesis treatment. Conventional breeding of this quantitative trait, however requires specific molecular markers. By means of biochemical experiments, we have established that the induced variation in linolenic acid content is associated with the fad3 gene encoding the microsomal 15 desaturase. Using a pair of primers specific to this gene and a doubled haploid progeny derived from a low linolenic x high linolenic acid F1hybrid, we have identified a polymorphism of the fad3 alleles between the low- and the high-linolenic acid genotypes. The structure exon/intron of the fad3 DNA sequence seems to be very similar to that of the Arabidopsis fad3 gene. The choice of the primer pair allows specific amplification of one of the two rapeseed fad3 genes. The value and contribution of specific markers to conventional plant breeding is discussed. 相似文献