With the form customization options in the form builder, you can easily change the outlook of your form and make it more unique and personalized. Disadvantages Whereas in-depth interviews are dynamic in nature, surveys are static, and don't allow for follow-up or further probing as to the "why" someone does something. What is Secondary Research? | Definition, Types, & Examples - Scribbr Conceptualization: J.Z., X.Y., A.C., L.L., Z.T. d Violin plots showing the Gene Set Variation Analysis (GSVA) scores of pathways in the five regions (T, n=12; M-T, n=21; M-P, n=21; P, n=10; LN, n=10) based on Stereo-seq data. The authors declare no competing interests. Lewis, S. M. et al. The cell components in each spot (bin50) in the slides were determined by SPOTlight26 using scRNA-seq data as the reference, which resulted in the spatial annotation of 9 main cell types (malignant cells and cholangiocytes were assigned as one main cell type; T cells and NK cells were also combined as one main cell type) (Fig. Investigation: L.W. S4e). To explore the mechanisms accounting for the high expression of SAAs in the Hep1 subtype, a Single-Cell Regulatory Network Inference and Clustering (SCENIC) analysis30 was applied and specific transcription factors (TFs), including FOSB, KLF6, NFKB1, STAT3, CEBPB, STAT2, and ETV6, were found to be more active in Hep1 compared with Hep2 based on the scRNA-seq data (Fig. Our forms have a unique QR code that you can add to your website or print in banners, business cards and the like. Pertea, M., Kim, D., Pertea, G. M., Leek, J. T. & Salzberg, S. L. Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown. Yuan, Y. Spatial heterogeneity in the tumor microenvironment. Secondary research maximizes the usefulness of data and unique specimens while minimizing risk to study volunteers since no new procedures are needed. Secondary research is also known as desk research since it involves synthesizing existing data that can be sourced from the internet. [9] Secondary research is an easy place to start when starting a new research project. 33, 495502 (2015). Our results showed that cells in the paratumor side (mainly hepatocytes) exhibited an increased inflammatory response and apoptosis levels, indicating that they had sustained severe damage. Dissecting and understanding the cancer ecosystem, especially that around the tumor margins, which have strong implications for tumor cell infiltration and invasion, are essential for exploring the mechanisms of tumor metastasis and developing effective new treatments. Margin area tissues (a 1cm-wide zone centered on the tumor border) were surgically removed from resected liver lobes from patients with liver cancer and were immersed in 90% Dulbeccos Modified Eagle medium (DMEM, Gibco, Gaithersburg, MD, USA) with 10% fetal bovine serum (FBS; Gibco), and were transported to the laboratory in a refrigerated container. Secondary Data - Types, Methods and Examples - Research Method = The only limitation is a dataset's format, structure, and whether or not it relates to the topic or problem at hand. Primary data is often used to answer specific research questions or to test hypotheses. 24, 235251 (2017). Wu, L., Yan, J., Bai, Y. et al. Margin areas were spatially divided into the tumor-adjacent part of margin areas (M-T) and the paratumor part of margin areas (M-P) according to the tumor border identified by pathologists based on adjacent H&E staining images. Grounded Theory Methods, Examples and Research Problem Examples, Types and Guide. Nat. Asterisks represent the significance levels of the performed tests (*P<0.05; **P<0.01; ***P<0.001). It can include quantitative and qualitative information. Primary research vs secondary research: Primary data is collected directly by the researcher (e.g., through interviews or experiments), while secondary data has already been collected by someone else (e.g., in government surveys or scientific publications). It can be recognized as a virtual aggregation of all secondary data outcomes. In some way, the internet is a virtual aggregation for all other sources of secondary research data. Students t-test was used for the analysis in panel c, a hypergeometric test was used in panels d, j and the Wilcoxon test was used in panel k. *P<0.05; **P<0.01; ***P<0.001. Can J. Gastroenterol. The increased expression levels of SAA1 and SAA2 in margin areas were significantly associated with a worse overall survival (OS) in ICC patients from validation cohort 2 (P<0.05 for SAA1 and P<0.05 for SAA2; Supplementary information, Fig. Intratumor heterogeneity and branched evolution revealed by multiregion sequencing. Research Data - Types Methods and Examples - Research Method 236, 53735386 (2021). Primary murine hepatocytes were treated with mouse recombinant GCP-2/CXCL6 protein (#ab9925, Abcam; 0ng/mL, 100ng/mL, 500ng/mL, and 1000ng/mL) for 36h. Monocytes were purified from the whole blood of healthy blood donors following a protocol approved by the Institutional Review Board at Fudan University, Zhongshan Hospital. The potential composition of each spot was pruned and renormalized using the top four cell types with respective probabilities in descending order, and the primary cell type was assigned for visualization. Grunwald, B. T. et al. When THP-1 reached confluence, the medium was supplemented with SAAs (#ab50232, Abcam) with a concentration of 500ng/mL or SAAs of 500ng/mL combinding with TLR2 inhibitor TLR2-IN-C29 (#S6597, Selleck, Shanghai, China) of 20M or CXCR2 inhibitor for 12h. Collected THP-1 cells were stored in cell staining buffer (#420201, Biolegend, San Diego, CA, USA) and incubated with Zombie NIR Fixable Viability Kit (#423106, Biolegend), PE anti-human CD45 antibody (#368510, Biolegend) and PE/Cyanine7 anti-human CD86 antibody (#374210, Biolegend). M-T, tumor side of the margin; M-P, paratumor side of the margin. 5. If you wish, you can share your forms QR code or embed it on your organizations website for easy access. We observed significant enrichment of immune cells and fibroblasts on the tumor side of the border compared with the paratumor side (Fig. Research data refers to any information or evidence gathered through systematic investigation or experimentation to support or refute a hypothesis or answer a research question. But, there are ways a researcher can use to improve the reliability and credibility of the secondary data in research. All our related research and data - Our World in Data Tanay, A. Components, Process and Tools. The liver cancer immune microenvironment: therapeutic implications for hepatocellular carcinoma. Blood 119, 18101820 (2012). Capture chips were generated following the Stereo-seq protocol.20 In brief, to generate the DNB array for in situ RNA capture, we first synthesized random 25-nucleotide coordinate identity (CID) -containing oligonucleotides, circularized with T4 DNA ligase, and splint oligonucleotides. After integrating Stereo-seq data with scRNA-seq, constructing a tumor border scanning and digitization model, and conducting additional bioinformatic analyses, we detected a high degree of cellular and transcriptional heterogeneities in a 500m-wide invasive zone, which is defined as the region within 250m on both sides of the tumor border. Our study did have some limitations. Chen, W. et al. Compared with data generated using other spatial transcriptome methods, our Stereo-seq data had unprecedented nanoscale resolution (diameter 220nm/spot) and expandable detection areas (10mm10mm), enabling a more precise view of all cell types and cellcell communications in the target area, as well as expanded spatial transcriptome analyses.1,9,20,55,56,57 Here, we characterized the transcriptional architecture in pseudo-spots using 25m25m squares (5050 bins/spot, bin50), representing approximately one cell. Scatter diagram showing the expression of SAAs in Hep1 (middle panel), and the expression of SAAs receptors in macrophages (right panel) on the LC5-M slide. Front. *P<0.05; **P<0.01; ***P<0.001. Immune cells were especially enriched within the first layer (0250m) closest to the border from the tumor side, where the immune cell fraction comprised more than 30% of all cell components (Fig. The commonly upregulated genes were also enriched in complement and coagulation cascades, apoptosis, and cholesterol metabolism, revealing an innate-immune related response and hepatocyte injury in Hep1 (Fig. Comprehensively defining the local immune ecosystem in the invasive zone would also advance our understanding of the mechanisms underlying tumor metastasis and might facilitate the development of more effective therapeutic strategies for other solid tumors. This is typically far cheaper and faster than primary research that requires experiments or analysis of raw data. There are two main types of market research - primary and secondary. 23, 17251755 (2016). The read structure was paired-end with Read 1, covering 30 bases inclusive of the 10bp cell barcode 1, 10bp cell barcode 2, and 10bp unique molecular identifier, and Read 2 containing 100 bases of the transcript sequence, and a 10bp sample index. Formplus allows you to easily add your form with respondents with the click of a button. Zheng, S. et al. While pre-registration can help to protect against researcher biases . 633, 185196 (2010). Your email address will not be published. Gene polymorphism of MUC15, MMP14, BRAF, and COL1A1 is associated with capsule formation in hepatocellular carcinoma. Amplified cDNA was purified using 60L of AMPure XP beads. Think of a library as an information storehouse that contains an aggregation of important information that can serve as valid data in different research contexts. The data generated in this study have been filed in the China Human Genetic Resources Service Management System (Record Number: 2022BAT2001), which is open to the public. Secondary data is called secondary because it is already manipulated. Google Scholar. The increased SAAs expression levels in the paratumor tissues in validation cohort 3 were also significantly correlated with a shorter OS (P<0.05 for OS; Supplementary information, Fig. The parameter condition was min.pct=0.1, logfc.threshold=0.15. Online Data Online data is data that is gathered via the internet. Published data is the most reliable secondary source of information. Secondary Articles", "LibGuides: Library Basics: Primary and Secondary Sources", "Diffusion of innovations in service organizations: systematic review and recommendations", https://www.thebalancesmb.com/what-is-secondary-market-research-2296890, "Introduction to primary research: observations, surveys, and interviews", https://www.geopoll.com/blog/primary-vs-secondary-research/, https://en.wikipedia.org/w/index.php?title=Secondary_research&oldid=1132940647, Creative Commons Attribution-ShareAlike License 4.0, This page was last edited on 11 January 2023, at 11:30. Below, we'll review the definition of secondary data, how it can be used by researchers, and the pros and cons of this type of research. Sometimes, secondary data is the only option because primary data is not available or difficult to reach. We identified two hepatocyte subtypes (Hep1 and Hep2), where Hep1 exhibited higher expression levels of SAA1 and SAA2 (Fig. S3e). Indexed sequencing libraries were constructed using the reagents provided in the C4 scRNA-seq kit as follows: (1) post-fragmentation size selection with AMPure XP beads; (2) end repair and A-tailing; (3) adapter ligation; (4) post-ligation purification with AMPure XP beads; (5) sample index PCR and size selection with AMPure XP beads. Finally, the value of the correlation coefficient was used to perform hierarchical clustering on all regions to elucidate the pattern. Briefly, monocytes were isolated from whole blood by positive sorting using anti-CD14-conjugated magnetic microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). From real-world insights to a countrys economic trends and information on the political climate, these sources are evolving into products that businesses can subscribe to by the year or pay for by-the-minute and on-demand projects. Typically, researchers donate several copies of dissertations to public and private libraries; especially in cases of academic research. Secondary research methods can also be categorized into qualitative and quantitative data collection methods. Anat. Newspaper are an excellent source of data collection historical research. Med. Serum amyloid A is a novel prognostic biomarker in hepatocellular carcinoma. 3f; Materials and methods). Pathways related to the epithelial-mesenchymal transition (EMT), the TMEs (e.g., hypoxia, angiogenesis, TGF- signaling), inflammation and apoptosis (e.g., apoptosis, complement, inflammatory response, reactive oxygen species), glycolysis and oxidative phosphorylation, and immune response (e.g., TNF- signaling via NF-B) were highly enriched in the tumor-adjacent part of margin areas compared to tumor tissues (Fig. d Representative images of the low and high SAAs/ARG1 ratio patients, and OS curves of 27 patients with primary liver cancer from validation cohort 4 grouped by the SAAs/ARG1 ratio determined using multiplexed IF staining (ARG1, CK19, SAAs, and DAPI). We found that Pattern 3 was significantly enriched in patients with tumor recurrence (P<0.001) compared to the other patterns, implying that macrophages in the invasive zone might be critical for tumor progression (Fig. The tissues were then fixed in methanol and incubated at 20C for 40min. We further validated the higher expression level of STAT3 in SAAs+ hepatocytes (Hep1) in the invasive zone by multiplexed IF staining (Fig. The higher expression of acute-phase protein genes, including SAA1, SAA2, Hemopexin (HPX), Orosomucoid 1 (ORM1), Orosomucoid 2 (ORM2), and several mediators of inflammation including Apolipoprotein A5 (APOA5) and Complement C3 (C3) in Hep1 indicated the damaged and inflammatory status of these cells (Supplementary information, Fig. While this method simplifies the data gathering process, the researcher must take care to depend solely on authentic sites when collecting information. 2b). All results were shown as the percentages of each tile, and statistical analyses were conducted to assess the heterogeneity in both the tangential and normal directions of the leading edge. (2023)Cite this article. [9] If possible, secondary research should be done before primary research, to determine what information is not already available. Nat. We also observed exhausted T cells scattered in this area, with a five-fold increase (from 0.09% to 0.54%) in the fraction of all cells from the third layer to the first layer, indicating an increase in T cell exhaustion as the distance from the border decreased (Supplementary information, Fig. S6e). Secondary data is the data that have been already collected for another purpose but has some relevance to your current research needs. "Search Smart: Types of Research: Secondary Research", "LibGuides: Scholarly Literature Types: Primary vs. Lancet 394, 11451158 (2019). While secondary research can be cost-effective and time-efficient, it requires the researcher to take extra care in ensuring that the data is authentic and valid. DeVault, G. (2019, June 25). After identifying the purpose of the research, the next thing to do is outline the research methodology. The normalized expression levels of different samples were converted to fragments per kb of transcript per million mapped (FPKM) fragments. There are generally four types of research data: This type of data involves the collection and analysis of numerical data. 10, 224 (2019). Adv. To systematically characterize the complex transcriptional architecture of human liver cancer, we processed fresh or freshly frozen T and P, M, and normal or metastatic LN using Stereo-seq with high resolution (diameter of 220nm/spot) and expandable areas (10mm10mm), and also analyzed some samples by scRNA-seq (discovery cohort, Fig. Spatially, immune cells such as macrophages were observed to accumulate primarily around the tumor border in margin areas, indicating a distinct immune microenvironment around the border (Fig. 15, 14841506 (2020). The macrophages in the invasive zone exhibited an M2-like phenotype with increased expression of CD163, MRC1, and SAAs receptor TLR2, whereas macrophages in the tumor tissue exhibited features of ECM remodeling and increased expression of MMP9 and MMP14 (Fig. In recent times, this method has become popular because the internet provides a large pool of both free and paid research resources that can be easily accessed with the click of a button. It has been widely used to characterize several types of solid tumors and the associated TMEs.11,12 However, scRNA-seq alone cannot provide spatial information.13 Further, due to the lack of multi-region sampling, intratumoral spatial heterogeneities at the single-cell resolution remain poorly understood.14,15,16 Recently, we developed SpaTial Enhanced REsolution Omics-sequencing (Stereo-seq), providing nanoscale resolution (diameter, 220nm/spot), expandable detection areas (10mm10mm), and the capacity to capture a few hundred spots of data per cell by combining DNA nanoball (DNB) patterned array chips and RNA in situ hybridization.17,18,19,20 Stereo-seq enables the in-depth characterization of functional and positional information for entire tumor ecosystems at the single-cell level, and precisely uncovers the cell composition, distribution, and cellcell communications in the TME, especially in the tumor margin areas. a The cellcell interactions between hepatocytes and other cell types indicated by the scRNA-seq data. The cell compositions and spatial distributions were highly heterogeneous in the four regions (T, M, P, and LN), with T/NK cells, B cells and fibroblasts being the most abundant cells aside from the predominant malignant cells and hepatocytes (Fig. What the secondary researcher does is to study already established patterns in previous researches and apply this information to the specific research context. An analysis of the ligandreceptor interaction using scRNA-seq data further suggested that the hepatocytes and macrophages in the invasive zone could also interactvia SAAs-TLR2 interactions (Fig. 22, 3848 (2020). Cold Spring Harb. Ji, A. L. et al. Advantages Surveys are a great way to collect significant amounts of representative quantitative data via primary research methods. [2][3], When conducting secondary research, authors may draw data from published academic papers, government documents, statistical databases, and historical records. Biol. You can also share your form to your social media pages using the social media direct sharing buttons available in the form builder. h Hierarchical clustering showing the transcriptional heterogeneity along the border and illustrating features of cell composition patterns acquired from a total of 2912 equally-divided subregions from the invasive zone in tissues from 16 patients with liver cancer.
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