IntroductionDifferentiating whether plant products are natural or artificial is of great importance in many practical fields, including forensic science, food safety, cosmetics, and fast-moving consumer goods. Information about the topographic distribution of compounds is an important criterion for answering this question. However, of equal importance is the likelihood that topographic spatial distribution information may provide important and valuable information for molecular mechanism study.MethodsIn this study, we took mescaline, a substance with hallucinogenic properties in cacti of the species Trichocereus pachanoi and Lophophora williamsii, as an example to characterize the spatial distribution of mescaline in plants and flowers by liquid chromatograph-mass spectrometry–matrix-assisted laser desorption/ionization mass spectrometry imaging at the macroscopic, tissue structure, and even cellular levels.ResultsAccording to our results, the distribution of mescaline in natural plant was concentrated on the active meristems, epidermal tissues, and protruding parts of Trichocereus pachanoi and Lophophora williamsii, while artificially spiked Lophophora diffusa products showed no such difference in their topographic spatial distribution.DiscussionThis difference in distribution pattern allowed us to distinguish between flowers that could synthesize mescaline on their own and those that had been artificially spiked with mescaline. The interesting topographic spatial distribution results, such as the overlap of the mescaline distribution map and micrographs of the vascular bundles, were consistent with the synthesis and transport theory of mescaline, indicating the potential for applying matrix-assisted laser desorption/ionization mass spectrometry imaging in botanical research.
How to sample a seizure plant:the role of visualization spatial distribution analysis of Lophophora williamsii as an exampleAbstract: Natural compounds in plants are often unevenly distributed, and determining the best sampling locations to obtain the most representative results is technically challenging. Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) can provide the basis for formulating sampling guideline. For a succulent plant sample, ensuring the authenticity and in situ nature of the spatial distribution analysis results during MSI analysis also needs to be thoroughly considered. In this study, we developed a well-established and reliable MALDI-MSI method based on preservation methods, slice conditions, auxiliary matrices, and MALDI parameters to detect and visualize the spatial distribution of mescaline in situ in L. williamsii. The MALDI-MSI results were validated using liquid chromatography–tandem mass spectrometry (LC–MS/MS). Low-temperature storage at −80°C and drying of ‘bookmarks’ were the appropriate storage methods for succulent plant samples and their flower samples, and cutting into 40μm thick sections at −20°C using gelatin as the embedding medium is the appropriate sectioning method. The use of DCTB (trans-2-[3-(4-tert-butylphenyl)-2-methyl-2-propenylidene]malononitrile) as an auxiliary matrix and a laser intensity of 45 are favorable MALDI parameter conditions for mescaline analysis. The region of interest (ROI) semi-quantitative analysis revealed that mescaline is concentrated in the epidermal tissues of L. williamsii as well as in the meristematic tissues of the crown. The study findings not only help to provide a basis for determining the best sampling locations for mescaline in L. williamsii, but they also provide a reference for the optimization of storage and preparation conditions for raw plant organs before MALDI detection.
Background: Previous randomized studies have assessed the possibility of omission of chemotherapy in some hormone receptor (HR)-positive and HER2-negative (HR+/HER2-) breast cancers (BC) based on gene pro ling test, e.g., Oncotype DX. The goal of this study was to evaluate if combination of six proliferation related biomarkers by immunohistochemistry (6-IHC) could be a cost-effective option in determining the necessity of adjuvant chemotherapy in HR+/HER2-BC.Methods: A retrospective analysis of HR+/HER2-BC patients was conducted in the First A liated Hospital of China Medical University from 2010 to 2016. The expression of 6 BC-related proliferation and invasion genes (Cathepsin L2, MMP11, CyclinB1, Aurora A, Survivin and Ki67) from Oncotype DX were analyzed through IHC (designated as 6-IHC). All the included patients were divided randomly at a 7:3 ratio into training and testing cohorts. The cutoff prognosis index (PI) of 6-IHC was determined by multivariate Cox risk regression analysis after calculating the PI of each patient in training cohort and con rmed in testing cohort. The patients were classi ed into "Low" and "High" risk groups based on the PI value.Kaplan-Meier (KM) method was used to analyze Disease-free survival (DFS) and overall survival (OS). 6-IHC score and other factors associated with survival bene t of adjuvant chemotherapy were compared with Ki67 index.Results: A total of 330 patients were included and divided into training cohort (n = 231) and validation cohort (n = 99). The receiver operating characteristic (ROC) curve analysis showed that the patients can be divided into 6-IHC score "High" and "Low" risk groups using the cut-off PI of 2.16. The 8-year DFS and OS were 54.6% and 69.2%, respectively in the 6-IHC score "High" risk group; 85.5% and 92.5%, respectively in the 6-IHC score "Low" risk group. The 8-year DFS and OS were 70.8% and 80.9%, respectively in the Ki67 "High" risk group, 77.7% and 87.6%, respectively in the Ki67 "Low" risk group. The KM curves showed that chemotherapy did not signi cantly improve the DFS in the 6-IHC score "Low" risk group (p = 0.830), but signi cantly improved the DFS in the 6-IHC score "High" risk group (P = 0.012).Conclusions: Combined 6-IHC score could be a reliable tool in predicting cancer-speci c recurrences and survival in HR+/HER2-BC patients and identifying patients who could bene t from adjuvant chemotherapy regardless of the involvement of axillary lymph node (ALN).
Background: Previous randomized studies have assessed the possibility of omission of chemotherapy in some hormone receptor (HR)-positive and HER2-negative (HR+/HER2-) breast cancers (BC) based on gene profiling test, e.g., Oncotype DX. The goal of this study was to evaluate if combination of six proliferation related biomarkers by immunohistochemistry (6-IHC) could be a cost-effective option in determining the necessity of adjuvant chemotherapy in HR+/HER2- BC.Methods: A retrospective analysis of HR+/HER2- BC patients was conducted in the First Affiliated Hospital of China Medical University from 2010 to 2016. The expression of 6 BC-related proliferation and invasion genes (Cathepsin L2, MMP11, CyclinB1, Aurora A, Survivin and Ki67) from Oncotype DX were analyzed through IHC (designated as 6-IHC). All the included patients were divided randomly at a 7:3 ratio into training and testing cohorts. The cutoff prognosis index (PI) of 6-IHC was determined by multivariate Cox risk regression analysis after calculating the PI of each patient in training cohort and confirmed in testing cohort. The patients were classified into “Low” and “High” risk groups based on the PI value. Kaplan-Meier (KM) method was used to analyze Disease-free survival (DFS) and overall survival (OS). 6-IHC score and other factors associated with survival benefit of adjuvant chemotherapy were compared with Ki67 index.Results: A total of 330 patients were included and divided into training cohort (n = 231) and validation cohort (n = 99). The receiver operating characteristic (ROC) curve analysis showed that the patients can be divided into 6-IHC score “High” and “Low” risk groups using the cut-off PI of 2.16. The 8-year DFS and OS were 54.6% and 69.2%, respectively in the 6-IHC score “High” risk group; 85.5% and 92.5%, respectively in the 6-IHC score “Low” risk group. The 8-year DFS and OS were 70.8% and 80.9%, respectively in the Ki67 “High” risk group, 77.7% and 87.6%, respectively in the Ki67 “Low” risk group. The KM curves showed that chemotherapy did not significantly improve the DFS in the 6-IHC score “Low” risk group (p = 0.830), but significantly improved the DFS in the 6-IHC score “High” risk group (P = 0.012).Conclusions: Combined 6-IHC score could be a reliable tool in predicting cancer-specific recurrences and survival in HR+/HER2- BC patients and identifying patients who could benefit from adjuvant chemotherapy regardless of the involvement of axillary lymph node (ALN).
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