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Fecal Microbiome Does Not Represent Whole Gut Microbiome 粪便微生物组并不代表整个肠道的微生物组
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2023-01-17 DOI: 10.1155/2023/6868417
Ji-Seon Ahn, Enkhchimeg Lkhagva, Sunjun Jung, Hyeon-Jin Kim, Hea-Jong Chung, Seong-Tshool Hong
The current gut microbiome research relies on the fecal microbiome under the assumption that the fecal microbiome represents the microbiome of the entire gastrointestinal (GI) tract. However, there have been growing concerns about using feces as a proxy to study the gut microbiome. Here, we comprehensively analyzed the composition of microbiome and metabolites in the feces and at 14 different locations of GI tracts of genetically homogenous sibling pigs to evaluate the validity of using feces as a proxy to the whole gut microbiome. The composition of intestinal microbes constituting the gut microbiome at each intestinal content and feces and their metabolic compositions were thoroughly investigated through metagenome sequencing and an ultraperformance LC-MS/MS, respectively. The fluctuation in the composition of the microbiome in the stomach and the small intestine became stabilized from the large intestine to feces and was able to be categorized into 3 groups. The taxonomic α-diversities measured by ACE (abundance-based coverage estimator) richness and Shannon diversity indicated that the microbiome in the large intestine was much more diverse than those of the small intestine and feces. The highly independent intestinal microbes in the stomach and the small intestine became flourished in the large intestine and converged into a community with tightly connected networks. β-Diversity analyses by NMDS plots, PCA, and unsupervised hierarchical clustering all showed that the diversities of microbiome compositions were lowest in feces while highest in the large intestine. In accordance with fluctuation of the composition of gut microbiome along with the GI tract, the metabolic composition also completely differed in a location-specific manner along with the GI tract. Comparative analysis of the fecal microbiome and metabolites with those of the whole GI tract indicated that fecal microbiome is insufficient to represent the whole gut microbiome.
目前的肠道微生物组研究依赖于粪便微生物组,认为粪便微生物组代表了整个胃肠道的微生物组。然而,越来越多的人担心使用粪便作为研究肠道微生物组的代理。在这里,我们综合分析了遗传同质兄弟猪粪便和胃肠道14个不同位置的微生物组和代谢物的组成,以评估将粪便作为整个肠道微生物组的替代品的有效性。分别通过宏基因组测序和高性能LC-MS/MS对各肠内容物和粪便中构成肠道微生物组的肠道微生物组成及其代谢组成进行了深入研究。胃和小肠微生物组组成的波动从大肠到粪便趋于稳定,可分为3组。利用ACE(丰度覆盖估计器)丰富度和Shannon多样性测量的分类α-多样性表明,大肠微生物组的多样性远高于小肠和粪便。胃和小肠中高度独立的肠道微生物在大肠中蓬勃发展,并融合成一个紧密相连的网络群落。通过NMDS图、PCA和无监督分层聚类分析的β-多样性分析均显示,粪便中微生物组的多样性最低,而大肠中微生物组的多样性最高。肠道微生物组的组成随胃肠道的变化而变化,代谢组成也随胃肠道的变化而完全不同。粪便微生物组和代谢产物与全胃肠道的比较分析表明,粪便微生物组不足以代表全肠道微生物组。
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引用次数: 2
Application of Bioinformatics Tools for the Prediction of Helper MicroRNAs for Improvement of Oncolytic Virus Efficacy 应用生物信息学工具预测辅助microrna以提高溶瘤病毒的疗效
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-12-29 DOI: 10.1155/2022/5756131
Nasrin Rastegarvand, H. Soleimanjahi, E. Arefian, M. Pourkarim
Purpose. Oncolytic Reoviruses, as a self-limiting virus, can be used in cancer treatment, because they have the ability to replicate in tumor cells selectively and destroy them. Studies show that some immune response proteins may interfere with the virus life cycle. So, the main aim of this bioinformatic study is to check which microRNA is able to target some reovirus inhibitory proteins. Experimental Design. By use of online bioinformatics software, the microRNAs that could target inhibitory genes were selected. Then, other features like content ++ score and cell type were checked and finally the eligible microRNAs were determined. Results. After choosing 15 inhibitory proteins, analysis was performed and finally 37 microRNAs which could target inhibitory proteins in colorectal cell lines were selected. In the end, by investigation of web-based tools, just two microRNAs were finalized. Conclusions and Clinical Relevance. This bioinformatic study shows that microRNA-140 and microRNA-92a have the potential to target some inhibitory proteins which interfere with oncolytic Reovirus replication and it may help in the optimal use of this virus as a cancer treatment. Because selective reproduction of Reovirus in tumor cells, as a nonchemical therapy, can be a good way to overcome this disease with broad advantages.
意图溶癌呼肠孤病毒作为一种自限性病毒,可以用于癌症治疗,因为它们具有选择性地在肿瘤细胞中复制并破坏它们的能力。研究表明,一些免疫反应蛋白可能会干扰病毒的生命周期。因此,这项生物信息学研究的主要目的是检查哪种微小RNA能够靶向某些呼肠孤病毒抑制蛋白。实验设计。利用在线生物信息学软件,筛选出能靶向抑制基因的微小RNA。然后,检查其他特征,如内容++评分和细胞类型,最终确定合格的微小RNA。后果在筛选出15种抑制蛋白后,进行分析,最终筛选出37种可靶向结直肠癌细胞系中抑制蛋白的微小RNA。最后,通过对基于网络的工具的调查,只有两个微小RNA最终确定。结论和临床相关性。这项生物信息学研究表明,微小RNA-140和微小RNA-92a有可能靶向一些干扰溶瘤性呼肠弧病毒复制的抑制蛋白,它可能有助于将这种病毒作为癌症治疗的最佳用途。因为在肿瘤细胞中选择性繁殖呼肠孤病毒,作为一种非化学疗法,是克服这种疾病的好方法,具有广泛的优势。
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引用次数: 0
Status of COVID-19 Infection and Vaccination in People Aged 18 Years and over in a Town: A Cross-Sectional Study in Turkey 土耳其某城镇18岁及以上人群COVID-19感染和疫苗接种状况的横断面研究
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-12-16 DOI: 10.1155/2022/9996615
Volkan Medeni, İrem Medeni
Objective. The objective of this study was to evaluate the cases of COVID-19 infection in people >18 years old and to investigate attitudes against vaccination and vaccine hesitancy and the factors affecting them. Methods. The data collection form used in the study was prepared by the researchers by screening the relevant literature. People over the age of 18 who agreed to participate in the study were invited to the family health center, and a questionnaire was applied by face-to-face interview method. The application time of a survey was 5-10 minutes. A total of 229 people were reached. Percentage of reaching was 95,4%. Results. The mean age of the participants was 44 , 13 ± 14 , 18 years. 31,9% of the participants in the study had COVID-19 infections. 69,9% of those who had COVID-19 infection caught the disease in the last year, and 93,2% were followed at home. Among the people with whom the participants lived, 21,6% of those who had COVID-19 infection were hospitalized and 5,7% died. Of the participants who expressed their opinion, 32,1% think that COVID-19 vaccines have serious side effects. Postvaccine side effects developed in 32,9% of the participants who had the COVID-19 vaccine, and 11,0% had a postvaccine infection. Conclusion. The results of our study revealed that the COVID-19 vaccine was highly accepted among the adult population in Turkey during the pandemic period. However, through this study, we found concerns about the side effects of the vaccine, the lack of confidence in the vaccine ingredient, and the effectiveness of the vaccine.
客观的本研究的目的是评估18岁以上人群中的新冠肺炎感染病例,并调查对疫苗接种和疫苗犹豫的态度及其影响因素。方法。研究人员通过筛选相关文献,编制了研究中使用的数据收集表。同意参与研究的18岁以上的人被邀请到家庭健康中心,并采用面对面访谈的方法进行问卷调查。调查的申请时间为5-10分钟。共联系了229人。达到率为95.4%。后果参与者的平均年龄分别为44、13±14、18岁。31.9%的研究参与者感染了新冠肺炎。去年,69,9%的新冠肺炎感染者感染了这种疾病,93.2%的人在家接受了随访。在参与者居住的人群中,感染新冠肺炎的患者中有21.5%住院,5.7%死亡。在表达意见的参与者中,32.1%的人认为新冠肺炎疫苗有严重的副作用。在接种新冠肺炎疫苗的参与者中,32.9%的人出现了疫苗后副作用,11,0%的人发生了疫苗后感染。结论我们的研究结果表明,在大流行期间,新冠肺炎疫苗在土耳其成年人中得到了高度接受。然而,通过这项研究,我们发现了对疫苗副作用、对疫苗成分缺乏信心以及疫苗有效性的担忧。
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引用次数: 1
Severe Inflammation Caused by Coinfection of PCV2 and Glaesserella parasuis Is Associated with Pyroptosis via Noncanonical Inflammasome Pathway PCV2和副猪小绿杆菌共同感染引起的严重炎症通过非典型炎性体途径与焦亡相关
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-11-16 DOI: 10.1155/2022/7227099
Jiahui An, Chao Zhang, Jinshuang Cai, Yufeng Li
Coinfections of porcine circovirus type 2 (PCV2) and Glaesserella parasuis (G. parasuis) are widely existing in the swine industry worldwide. However, the mechanisms for this coinfection remain unclear. The aim of this study is to assess whether the coinfection PCV2 and G. parasuis would affect the inflammatory response and related mechanisms. In this study, BALB/c mice and RAW264.7 cells were used to study the inflammation and related mechanism caused by the coinfection of PCV2 and G. parasuis. Coinfection with PCV2 and G. parasuis significantly increased the mortality of mice and led to the development of more severe lung and spleen lesions compared with single agent infection. Especially, coinfection significantly increased the bacterial loads in the lungs. Coinfection with PCV2 and G. parasuis can enhance RAW264.7 cell phagocytosis and elimination to G. parasuis. Cell death rate of cells increased in coinfection was measured with Flow cytometry. Moreover, coinfection led to the downregulation of the expression of TNFα and IL-8 in comparison with G. parasuis infection, but the maturation of interleukin-1β (IL-1β) was significantly upregulated. Our study firstly revealed that coinfection of PCV2 and G. parasuis can increase the phagocytosis of cells to G. parasuis, and LPS in the cytoplasm will induce the maturation of caspase-11 and lead to the cleavage of Gasdermin D (GSDMD) to cause pyroptosis by noncanonical pathway. The revealing of mechanisms associated with coinfection with PCV2 and G. parasuis will provide a scientific basis for investigating the synergistic infection mechanisms between viruses and bacteria.
猪圆环病毒2型(PCV2)和副猪格雷瑟菌(G.副猪)的共感染在世界范围内广泛存在。然而,这种合并感染的机制尚不清楚。本研究的目的是评估PCV2和副猪螺旋体共同感染是否会影响炎症反应及其相关机制。本研究采用BALB/c小鼠和RAW264.7细胞,研究PCV2和副猪螺旋体共同感染引起的炎症及相关机制。与单药感染相比,PCV2和副猪G.联合感染可显著提高小鼠死亡率,并导致更严重的肺和脾脏病变。特别是,合并感染显著增加了肺部的细菌负荷。PCV2与副猪螺旋体共感染可增强RAW264.7细胞对副猪螺旋体的吞噬和清除。用流式细胞术检测合并感染增加的细胞死亡率。此外,与副猪螺旋体感染相比,共感染导致TNFα和IL-8的表达下调,但白细胞介素-1β (IL-1β)的成熟水平显著上调。本研究首次发现PCV2与副猪螺旋体共感染可增加细胞对副猪螺旋体的吞噬能力,细胞质中的LPS可诱导caspase-11成熟,导致Gasdermin D (GSDMD)裂解,通过非典型途径引起焦亡。揭示PCV2与副猪弧菌共感染的相关机制,将为进一步研究病毒与细菌的协同感染机制提供科学依据。
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引用次数: 0
Deletion of a Putative GPI-Anchored Protein-Encoding Gene Aog185 Impedes the Growth and Nematode-Trapping Efficiency of Arthrobotrys oligospora by Disrupting Transmembrane Transport Homeostasis gpi锚定蛋白编码基因Aog185的缺失通过破坏跨膜运输稳态来阻碍少孢子节虫的生长和捕获线虫的效率
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-11-14 DOI: 10.1155/2022/8738290
Hui Peng, Hengqian Lu, Xinyuan Dong, Xiao Liang, Kangliang Sheng, Jingmin Wang, X. Kong, X. Zha, Yongzhong Wang
Nematode-trapping fungus (NTF) is a crucial predator of nematodes, which can capture nematodes by developing specific trapping devices. However, there is limited understanding of the role and mechanism of cell surface proteins attached to the surface of mycelia or trapping cells. Here, the effects of a putative GPI-anchored protein-encoding gene Aog185 on the growth and nematode-trapping efficiency of A. oligospora were investigated. Compared to the wild-type (WT) strain, the ΔAog185 mutant grew more slowly, exhibited a 20% decrease in conidiation, delayed conidial germination, generated fewer traps, attenuated nematode trapping efficiency, and was more sensitive to chemical stressors. Transcriptomic analysis indicated that a large number of transmembrane transport-related genes were differentially expressed between the WT and ΔAog185 mutant strains. Aog185 deletion could damage the intrinsic components of the membrane and cytoskeleton. Specifically, knockout of Aog185 disrupted transmembrane transport homeostasis during the phagocytosis, cell autophagy, and oxidative phosphorylation processes, which were associated with the fusion of cells and organelle membranes, transport of ions and substrates, and energy metabolism. Hence, the putative GPI-anchored protein-encoding gene Aog185 may contribute to the lifestyle switch of NTF and nematode capture, and the effect of Aog185 gene on cell transmembrane transport is considered key to this process. Our findings provide new insights into the mechanism of Aog185 gene during the process of nematode trapping by NTF.
诱捕线虫真菌(Nematode-trapping fungus, NTF)是一种重要的线虫捕食者,它可以通过形成特定的诱捕装置捕获线虫。然而,对附着在菌丝或诱捕细胞表面的细胞表面蛋白的作用和机制了解有限。本文研究了推定的gpi锚定蛋白编码基因Aog185对寡孢单胞菌生长和捕获线虫效率的影响。与野生型(WT)菌株相比,ΔAog185突变体生长更慢,分生孢子减少20%,分生孢子萌发延迟,产生的陷阱更少,捕获线虫的效率降低,对化学胁迫更敏感。转录组学分析表明,WT和ΔAog185突变株之间存在大量跨膜运输相关基因的差异表达。Aog185缺失会破坏细胞膜和细胞骨架的固有成分。具体来说,敲除Aog185破坏了吞噬、细胞自噬和氧化磷酸化过程中的跨膜运输稳态,这些过程与细胞和细胞器膜的融合、离子和底物的运输以及能量代谢有关。因此,假定的gpi锚定蛋白编码基因Aog185可能有助于NTF和线虫捕获的生活方式转换,并且Aog185基因对细胞跨膜运输的影响被认为是这一过程的关键。本研究为研究Aog185基因在NTF诱捕线虫过程中的作用机制提供了新的思路。
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引用次数: 0
Similarities and Differences among Species Closely Related to Candida albicans: C. tropicalis, C. dubliniensis, and C. auris 与白色念珠菌密切相关的物种的异同:热带念珠菌、都柏林念珠菌和耳念珠菌
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-11-09 DOI: 10.1155/2022/2599136
Dorota Satala, Magdalena Juszczak, Ewelina Wronowska, Magdalena Surowiec, Kamila Kulig, Andrzej Kozik, Maria Rapala-Kozik, Justyna Karkowska-Kuleta
Although Candida species are widespread commensals of the microflora of healthy individuals, they are also among the most important human fungal pathogens that under certain conditions can cause diseases (candidiases) of varying severity ranging from mild superficial infections of the mucous membranes to life-threatening systemic infections. So far, the vast majority of research aimed at understanding the molecular basis of pathogenesis has been focused on the most common species—Candida albicans. Meanwhile, other closely related species belonging to the CTG clade, namely, Candida tropicalis and Candida dubliniensis, are becoming more important in clinical practice, as well as a relatively newly identified species, Candida auris. Despite the close relationship of these microorganisms, it seems that in the course of evolution, they have developed distinct biochemical, metabolic, and physiological adaptations, which they use to fit to commensal niches and achieve full virulence. Therefore, in this review, we describe the current knowledge on C. tropicalis, C. dubliniensis, and C. auris virulence factors, the formation of a mixed species biofilm and mutual communication, the environmental stress response and related changes in fungal cell metabolism, and the effect of pathogens on host defense response and susceptibility to antifungal agents used, highlighting differences with respect to C. albicans. Special attention is paid to common diagnostic problems resulting from similarities between these species and the emergence of drug resistance mechanisms. Understanding the different strategies to achieve virulence, used by important opportunistic pathogens of the genus Candida, is essential for proper diagnosis and treatment.
虽然念珠菌是健康个体微生物群的广泛共生体,但它们也是最重要的人类真菌病原体之一,在某些条件下可引起不同严重程度的疾病(念珠菌病),从轻微的粘膜浅表感染到危及生命的全身感染。到目前为止,绝大多数旨在了解发病机制的分子基础的研究都集中在最常见的物种-白色念珠菌上。与此同时,其他与CTG分支密切相关的种,即热带念珠菌(Candida tropical)和都柏林念珠菌(Candida dubliniensis)在临床实践中越来越重要,还有一个相对较新发现的种——耳念珠菌(Candida auris)。尽管这些微生物之间的关系密切,但在进化过程中,它们似乎已经发展出不同的生化、代谢和生理适应,它们利用这些适应来适应共生生态位并实现完全的毒力。因此,本文综述了热带镰刀菌、都柏林镰刀菌和耳镰刀菌的毒力因子、混合物种生物膜的形成和相互通讯、环境应激反应和真菌细胞代谢的相关变化、病原体对宿主防御反应和抗真菌药物敏感性的影响,并重点介绍了白色镰刀菌的不同。特别注意由于这些物种之间的相似性和耐药性机制的出现而引起的常见诊断问题。了解念珠菌属重要的机会致病菌实现毒力的不同策略,对于正确诊断和治疗至关重要。
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引用次数: 0
Biomolecules of the Horseshoe Crab’s Hemolymph: Components of an Ancient Defensive Mechanism and Its Impact on the Pharmaceutical and Biomedical Industry 马蹄蟹血淋巴的生物分子:一种古老防御机制的组成部分及其对制药和生物医学工业的影响
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-10-27 DOI: 10.1155/2022/3381162
M. Ashrafuzzaman, M. H. Razu, N. Showva, T. A. Bondhon, M. Moniruzzaman, Sad Al Rezwan Rahman, Md. Raisul Islam Rabby, F. Akter, Mala Khan
Without adaptive immunity, invertebrates have evolved innate immune systems that react to antigens on the surfaces of pathogens. These defense mechanisms are included in horseshoe crab hemocytes’ cellular responses to pathogens. Secretory granules, large (L) and small (S), are found on hemocytes. Once the invasion of pathogens is present, these granules release their contents through exocytosis. Recent data in biochemistry and immunology on the granular constituents of granule-specific proteins are stored in large and small granules which are involved in the cell-mediated immune response. L-granules contain most clotting proteins, which are necessary for hemolymph coagulation. They also include tachylectins; protease inhibitors, such as cystatin and serpins; and anti-lipopolysaccharide (LPS) factors, which bind to LPS and agglutinate bacteria. Big defensin, tachycitin, tachystatin, and tachyplesins are some of the essential cysteine-rich proteins in S-granules. These granules also contain tachycitin and tachystatins, which can agglutinate bacteria. These proteins in granules and hemolymph act synergistically to fight infections. These biomolecules are antimicrobial and antibacterial, enabling them to be drug resistant. This review is aimed at explaining the biomolecules identified in the horseshoe crab’s hemolymph and their application scopes in the pharmaceutical and biotechnology sectors.
在没有适应性免疫的情况下,无脊椎动物进化出了对病原体表面抗原做出反应的先天免疫系统。这些防御机制包含在鲎血细胞对病原体的细胞反应中。血细胞上有分泌颗粒,大(L)和小(S)。一旦病原体入侵,这些颗粒就会通过胞吐作用释放其内含物。生物化学和免疫学中关于颗粒特异性蛋白质颗粒成分的最新数据存储在参与细胞介导的免疫反应的大颗粒和小颗粒中。L-颗粒含有大多数凝血蛋白,这些蛋白是血淋巴凝固所必需的。它们还包括快速凝集素;蛋白酶抑制剂,如胱抑素和丝氨酸蛋白酶抑制剂;以及与LPS结合并凝集细菌的抗脂多糖(LPS)因子。大防御素、速激肽、速抑制素和速激肽是S颗粒中一些必需的富含半胱氨酸的蛋白质。这些颗粒还含有速激肽和速激他汀类药物,它们可以凝集细菌。颗粒和血淋巴中的这些蛋白质协同作用对抗感染。这些生物分子具有抗菌性和抗菌性,使它们具有耐药性。本文旨在解释鲎血淋巴中鉴定的生物分子及其在制药和生物技术领域的应用范围。
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引用次数: 2
Pasteurella multocida Toxin Aggravates Ligatured-Induced Periodontal Bone Loss and Inflammation via NOD-Like Receptor Protein 3 Inflammasome 多杀性巴氏杆菌毒素通过nod样受体蛋白3炎性体加重结扎诱导的牙周骨丢失和炎症
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-10-04 DOI: 10.1155/2022/3305695
Yineng Han, P. Gao, Qiaolin Yang, Yiping Huang, L. Jia, Yunfei Zheng, Weiran Li
NOD-like receptor family pyrin domain-containing 3 (NLRP3) inflammasome is reportedly involved in periodontal pathogenesis. Pasteurella multocida toxin (PMT) is the major virulence factor of Pasteurella multocida strains, which belongs to the nonoral gram-negative facultative rods (GNFR). The existence of GNFR and their toxin may aggravate periodontitis. Therefore, it is important to unclose the regulatory mechanisms of PMT in periodontitis. However, the involvement of NLRP3 inflammasome and PMT in periodontitis remain unclear. The results showed that NLRP3 expression was increased in periodontitis mice by immunohistochemical staining and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Nlrp3-/- mice showed less periodontal bone loss and lower abundances of Pasteurella multocida by 16S rRNA sequencing. PMT promoted NLRP3 expressions by activating nuclear factor kappa light chain enhancer of B cells (NF-κB) pathway and activated NLRP3 inflammasome. This effect was reversed by NLRP3 inhibitor MCC950. Furthermore, PMT aggravated periodontal bone loss and inflammation in WT mice, while MCC950 attenuated periodontal bone loss and inflammation. The Nlrp3-/- periodontitis models with PMT local injection showed less bone loss and inflammation compared with WT periodontitis mice after PMT treatment. Taken together, our results showed that PMT aggravates periodontal response to the ligature by promoting NLRP3 expression and activating NLRP3 inflammasome, suggesting that NLRP3 may be an effective target for the treatment of periodontitis caused by GNFR and MCC950 may be a potential drug against this disease.
据报道,nod样受体家族pyrin结构域- 3 (NLRP3)炎症小体参与牙周发病。多杀性巴氏杆菌毒素(PMT)是多杀性巴氏杆菌属非口服革兰氏阴性兼性杆状菌(GNFR)的主要毒力因子。GNFR及其毒素的存在可加重牙周炎。因此,揭示PMT在牙周炎中的调节机制具有重要意义。然而,NLRP3炎性体和PMT在牙周炎中的作用尚不清楚。免疫组化染色和定量逆转录聚合酶链反应(qRT-PCR)结果显示,牙周炎小鼠NLRP3表达升高。通过16S rRNA测序,Nlrp3-/-小鼠牙周骨丢失较少,多杀性巴氏杆菌丰度较低。PMT通过激活B细胞核因子κB轻链增强子(NF-κB)通路,激活NLRP3炎性体,促进NLRP3表达。这种作用被NLRP3抑制剂MCC950逆转。此外,PMT加重了WT小鼠的牙周骨质流失和炎症,而MCC950则减轻了牙周骨质流失和炎症。局部注射PMT的Nlrp3-/-牙周炎模型与PMT治疗后的WT牙周炎小鼠相比,骨质流失和炎症反应较少。综上所述,我们的研究结果表明,PMT通过促进NLRP3表达和激活NLRP3炎性体,加重牙周对结膜的反应,提示NLRP3可能是治疗GNFR引起的牙周炎的有效靶点,MCC950可能是治疗该疾病的潜在药物。
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引用次数: 0
Correlation between HPV PCNA, p16, and p21 Expression in Lung Cancer Patients 肺癌患者HPV PCNA、p16和p21表达的相关性
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-09-28 DOI: 10.1155/2022/9144334
B. Marcos, T. H. A. de Oliveira, C. M. D. do Amaral, M. T. Muniz, A. C. Freitas
Purpose. Evaluate if human papillomavirus (HPV) infection in lung cancer patients might be helping cancer development by altering p16, p21, and PCNA, key human genes involved in cell proliferation and tumor development. Methods. 63 fresh-frozen (FF) and formalin-fixed paraffin-embedded (FFPE) samples from lung tumor patients were used to detect HPV by PCR, followed by genotype through sequencing. The host gene expressions of p21, p16, and PCNA were quantified by qPCR in both FF and FFPE samples, and the expression of viral oncogenes E5, E6, and E7 was also measured by qPCR in 19 FF samples. Results. 74.6% of samples were positive for HPV, 33/44 FFPE samples and 14/19 FF samples. HPV-16 and HPV-18 were detected in 31/33 and 7/33 FFPE, respectively, and HPV-16 was the only type in FF samples. E5, E6, and E7 were expressed in 10/19, 2/19, and 4/19 FF samples, respectively. The p16 RNAm expression was higher in FF HPV+ samples and FFPE+FF HPV+ samples, while p21 showed higher expression in all HPV- samples. In turn, the PCNA expression was higher in HPV+ FF samples; however, in FFPE and FFPE+FF samples, PCNA was higher in HPV- samples. In FF samples, PCNA, p16, and p21 showed a significant positive correlation as well as E5 and E7, and E5 was inversely correlated to p21. In FFPE, also, a positive correlation was observed between PCNA HPV+ and p21 HPV+ and PCNA HPV+ and p16 HPV. In FF+FFPE analysis, a direct correlation was found between PCNA HPV+ and p21 HPV+, p21 HPV+ and p16 HPV+, and PCNA HPV- and p16 HPV-, and an inverse correlation between PCNA HPV+ and p16 HPV+. Also, the p16 protein was positive in 10 HPV+ samples and 1 HPV-. Conclusions. Our data show that lung cancer patients from Northeast Brazil have a high prevalence of HPV, and the virus also expresses its oncogenes and correlates with key human genes involved in tumor development. This data could instigate the development of studies focused on preventive strategies, such as vaccination, used as a prognostic indicator and/or individualized therapy.
意图评估癌症患者的人乳头瘤病毒(HPV)感染是否可能通过改变p16、p21和PCNA(参与细胞增殖和肿瘤发展的关键人类基因)来帮助癌症的发展。方法。采用63份新鲜冷冻(FF)和福尔马林固定石蜡包埋(FFPE)肺肿瘤患者的样本,通过PCR检测HPV,然后通过测序检测基因型。通过qPCR对FF和FFPE样本中p21、p16和PCNA的宿主基因表达进行定量,并通过qPCR测定19个FF样本中病毒致癌基因E5、E6和E7的表达。后果74.6%的样本为HPV阳性,33/44份FFPE样本和14/19份FF样本。在31/33和7/33的FFPE中分别检测到HPV-16和HPV-18,并且HPV-16是FF样品中唯一的类型。E5、E6和E7分别在10/19、2/19和4/19 FF样品中表达。p16RNAm在FF HPV+样本和FFPE+FF HPV+样本中表达较高,而p21在所有HPV-样本中表达均较高。反过来,PCNA在HPV+FF样本中的表达更高;然而,在FFPE和FFPE+FF样本中,HPV样本中PCNA较高。在FF样本中,PCNA、p16和p21与E5和E7呈显著正相关,E5与p21呈负相关。在FFPE中,PCNA HPV+和p21 HPV+以及PCNA HPW+和p16 HPV之间也观察到正相关。在FF+FFPE分析中,发现PCNA HPV+与p21 HPV+、p21 HPW+与p16 HPV+以及PCNA HPV-与p16 HPV-之间存在直接相关性,而PCNA HPW+和p16 HPV-之间存在反相关性。此外,p16蛋白在10个HPV+样本和1个HPV-样本中呈阳性。结论。我们的数据显示,来自巴西东北部的癌症患者的HPV患病率很高,该病毒也表达其致癌基因,并与参与肿瘤发展的关键人类基因相关。这些数据可以推动专注于预防策略的研究的发展,如疫苗接种,用作预后指标和/或个体化治疗。
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引用次数: 1
lncRNA MANCR Inhibits NK Cell Killing Effect on Lung Adenocarcinoma by Targeting miRNA-30d-5p lncRNA MANCR通过靶向miRNA-30d-5p抑制NK细胞对肺腺癌的杀伤作用
IF 3.4 2区 生物学 Q1 Immunology and Microbiology Pub Date : 2022-09-16 DOI: 10.1155/2022/4928635
Yunping Lu, Xiao Rao, W. Zheng, Yinggan Du, Jianbo Xue, Kan Huang
Background. NK cells are imperative in spontaneous antitumor response of various cancers. Currently, lncRNAs are considered important modulators of the tumor microenvironment. This study investigated the molecular mechanism by which mitotically associated long noncoding RNA (MANCR) controls killing effect of NK cells on lung adenocarcinoma (LUAD) in the tumor microenvironment. Methods. The interplay between MANCR and miRNA-30d-5p was analyzed by bioinformatics. Expression of MANCR mRNA and miRNA-30d-5p was examined using qRT-PCR. Dual-luciferase reporter and RIP assays were utilized to verify the targeted relationship between MANCR and miRNA-30d-5p. To investigate regulation of MANCR/miRNA-30d-5p axis in NK cell killing effect on LUAD cells, western blot tested the protein level of perforin and granzyme B. ELISA determined the level of IFN-γ. CytoTox 96 Non-Radioactive Cytotoxicity Assay kit was applied for cytotoxicity detection of NK cells. Perforin and granzyme B fluorescence intensity was measured via immunofluorescence, and cell apoptosis levels were also revealed via flow cytometry. Results. MANCR was found to be upregulated, while miRNA-30d-5p expression was downregulated in LUAD tissues. Overexpression of MANCR in LUAD cells significantly reduced NK cell IFN-γ secretion, expression of granzyme B and perforin, and NK cell killing effect. In addition, MANCR could target and downregulate miRNA-30d-5p expression, and miRNA-30d-5p overexpression reversed the inhibition of NK cell killing effect caused by MANCR overexpression. Conclusion. MANCR inhibited the killing effect of NK cells on LUAD via targeting and downregulating miRNA-30d-5p and provided new ideas for antitumor therapy based on tumor microenvironment.
背景。NK细胞在各种肿瘤的自发抗肿瘤反应中起重要作用。目前,lncrna被认为是肿瘤微环境的重要调节剂。本研究探讨了有丝分裂相关长链非编码RNA (MANCR)在肿瘤微环境中调控NK细胞对肺腺癌(LUAD)杀伤作用的分子机制。方法。生物信息学分析了MANCR与miRNA-30d-5p之间的相互作用。采用qRT-PCR检测MANCR mRNA和miRNA-30d-5p的表达。利用双荧光素酶报告基因和RIP检测来验证MANCR与miRNA-30d-5p之间的靶向关系。为了研究MANCR/miRNA-30d-5p轴在NK细胞对LUAD细胞杀伤作用中的调控作用,western blot检测了穿孔素和颗粒酶b的蛋白水平,ELISA检测了IFN-γ水平。采用CytoTox 96非放射性细胞毒性检测试剂盒对NK细胞进行细胞毒性检测。免疫荧光法检测穿孔素和颗粒酶B荧光强度,流式细胞术检测细胞凋亡水平。结果。在LUAD组织中,MANCR表达上调,而miRNA-30d-5p表达下调。LUAD细胞过表达MANCR可显著降低NK细胞IFN-γ分泌、颗粒酶B和穿孔素的表达,降低NK细胞杀伤作用。此外,MANCR可以靶向并下调miRNA-30d-5p的表达,miRNA-30d-5p过表达逆转了MANCR过表达对NK细胞杀伤的抑制作用。结论。MANCR通过靶向下调miRNA-30d-5p抑制NK细胞对LUAD的杀伤作用,为基于肿瘤微环境的抗肿瘤治疗提供了新的思路。
{"title":"lncRNA MANCR Inhibits NK Cell Killing Effect on Lung Adenocarcinoma by Targeting miRNA-30d-5p","authors":"Yunping Lu, Xiao Rao, W. Zheng, Yinggan Du, Jianbo Xue, Kan Huang","doi":"10.1155/2022/4928635","DOIUrl":"https://doi.org/10.1155/2022/4928635","url":null,"abstract":"Background. NK cells are imperative in spontaneous antitumor response of various cancers. Currently, lncRNAs are considered important modulators of the tumor microenvironment. This study investigated the molecular mechanism by which mitotically associated long noncoding RNA (MANCR) controls killing effect of NK cells on lung adenocarcinoma (LUAD) in the tumor microenvironment. Methods. The interplay between MANCR and miRNA-30d-5p was analyzed by bioinformatics. Expression of MANCR mRNA and miRNA-30d-5p was examined using qRT-PCR. Dual-luciferase reporter and RIP assays were utilized to verify the targeted relationship between MANCR and miRNA-30d-5p. To investigate regulation of MANCR/miRNA-30d-5p axis in NK cell killing effect on LUAD cells, western blot tested the protein level of perforin and granzyme B. ELISA determined the level of IFN-γ. CytoTox 96 Non-Radioactive Cytotoxicity Assay kit was applied for cytotoxicity detection of NK cells. Perforin and granzyme B fluorescence intensity was measured via immunofluorescence, and cell apoptosis levels were also revealed via flow cytometry. Results. MANCR was found to be upregulated, while miRNA-30d-5p expression was downregulated in LUAD tissues. Overexpression of MANCR in LUAD cells significantly reduced NK cell IFN-γ secretion, expression of granzyme B and perforin, and NK cell killing effect. In addition, MANCR could target and downregulate miRNA-30d-5p expression, and miRNA-30d-5p overexpression reversed the inhibition of NK cell killing effect caused by MANCR overexpression. Conclusion. MANCR inhibited the killing effect of NK cells on LUAD via targeting and downregulating miRNA-30d-5p and provided new ideas for antitumor therapy based on tumor microenvironment.","PeriodicalId":9844,"journal":{"name":"Cellular Microbiology","volume":null,"pages":null},"PeriodicalIF":3.4,"publicationDate":"2022-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46310364","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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Cellular Microbiology
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