The secretory carrier-associated membrane proteins (SCAMPs) are associated with the development of multiple human cancers. The role of SCAMPs in acute myeloid leukemia (AML), however, remains to be identified. In the present study, we explored expression patterns and prognostic value of SCAMPs and network analysis of SCAMPs-related signaling pathways in AML using Oncomine, GEPIA, cBioPortal, LinkedOmics, DAVID and Metascape databases. Genetic alteration analysis revealed that the mutation rate of SCAMP genes was below 1% (9/1272) in AML, and there was no significant correlation between SCAMPs gene mutation and AML prognosis. However, the SCAMP2/5 mRNA levels were significantly higher in AML patients than in healthy controls. Moreover, high mRNA expressions of SCAMP2/4/5 were associated with poor overall survival, which might be due to that SCAMP2/4/5 and their co-expressed genes were associated with multiple pathways related to tumorigenesis and progression, including human T-cell leukemia virus 1 infection, acute myeloid leukemia, mTOR and NF-kappa B signaling pathways. These results suggest that SCAMP2/4/5 are potential prognostic markers for AML, and that SCAMP2 and SCAMP5 individually or in combination may be used as diagnostic markers for AML. SCAMPs control intracellular trafficking and signaling involved in cell–cell adhesion, cancer migration and invasion6. Based on the variable presence of multiple N-terminal asparagine-proline-phenylalanine (NPF) repeats, human SCAMPs are divided into two groups: SCAMP1/2/3 (with NPF repeats) and SCAMP4/5 (without NPF repeats), whereas, SCAMPs with the same NPF repeats may have distinct functions. Both groups have four central transmembrane regions (TMRs) and cytoplasmic tail. TMRs participate in membrane transport and traffic with their interacting partners on the membrane-cytosol interface8. Importantly, alterations in cytoskeletal pathways mediated by SCAMPs can affect cell–cell adhesion and may result in cell polarity loss and the increase of cell motility and invasion via changing components of the plasma membrane10. In consistent with these reports, dysregulation of SCAMPs has been found in series of human malignancies, such as hepatocellular carcinoma11, lung cancer12; breast cancer5, colorectal cancer2, ovarian cancer3, pancreatic cancer and gallbladder cancer13. Recently, the role of SCAMPs in human cancers has been a topic of increasing interest.The secretory carrier-associated membrane proteins (SCAMPs), a family of transcription factors encoded by five SCAMP genes in eukaryotes, are ubiquitously expressed in secretory membrane15. AML is the most common subtype in adult acute leukemia, accounting for 80% morbidity. It is estimated that there will be 20,240 new cases, and 11,400 deaths from AML in 2021 in USA16. Despite improvements in multi-agent chemotherapy, chemoimmunotherapy, targeted therapy and allogeneic stem cell transplantation for clinical management of leukemia, the 5-year relative survival was only 29.5% (2011–2017) due to chemotherapy resistance, immune rejection, and poor adherence to treatment20. Notably, current medical science lacks reliable and efficient prognostic biomarkers to enable early diagnosis and accurate prediction of prognosis for AML. Therefore, there is an urgent need to explore molecular biomarkers and therapeutic targets to enhance prognostic capabilities and to promote individualized treatment in the era of precision medicine.Acute myeloid leukemia (AML) is a malignant clonal disease of hematopoietic tissue characterized by dysregulated proliferation, impaired apoptosis, and disrupted blasts of myeloid lineage differentiation, accompanied by severe infections, anemia and haemorrhageUp to this point in time, the roles of SCAMPs in AML remain poorly understood. Herein, we comprehensively analyzed the relationships between the five SCAMPs and AML based on the data from Oncomine, GEPIA , cBioPortal, LinkedOmics and DAVID , as a means of assessing SCAMP expression patterns, potential functions, and prognostic utility in the context of AML.This study has been approved by the Ethics Committee of the First Affiliated Hospital of Xiamen University . The data was retrieved from published literature, and all analysis were performed in accordance with the Declaration of Helsinki.https://www.oncomine.org) database is an online tool to analyze, standardize, and process tumor microarray transcriptomic data. This database was used to analyze SCAMP expression profiles between AML samples and normal controls. The normal controls were data from bone marrow. P-values were generated through Student’s t-tests, and the cut-off criteria for Oncomine analyses in this study were a p-value of 0.01 and a fold-change value of 221.The Oncomine v4.5 (http://gepia.cancer-pku.cn) platform can be used to analyze differential expression profiles associated with various types of tumors in The Cancer Genome Atlas (TCGA) (http://tcga-data.nci.nih.gov/tcga/) and the Genotype-Tissue Expression Project (GTEx)  databases, incorporating RNA-seq expression data for 9,736 tumor samples and 8,587 control samples. Here GEPIA was used for distinct SCAMP isoforms expression in AML, and it was also used to observe the relationship between SCAMP expression and overall survival of patients with AML. Data from healthy whole blood samples were used as normal controls22.The GEPIA (http://www.linkedomics.org/admin.php) is a software tool used to disseminate data pertaining to all 32 cancer types included in the TCGA with a focus on interpreting and discovering attributes associations, thereby complementing other available databases. This database allows for analyses of multiomic datasets23. We conducted a co-expression analysis for SCAMPs using the LinkedOmics AML datasets.LinkedOmics  leverages the GEO (Gene Expression Omnibus) and TCGA databases, profiling mutations, copy-number alterations (CNAs) from GISTIC (Genomic Identification of Significant Targets in Cancer), and mRNA and protein expression Z-scores (RNASeq V2 RSEM and RPPA)25. Alteration of SCAMP genes status in AML patients was determined using the online cancer genomics cBioPortal.CBioPortal (https://david.ncifcrf.gov/) database27 was utilized to conduct Kyoto Encyclopedia of Genes and Genomes (KEGG) and gene ontology (GO) analyses of SCAMPs. Moreover, GO enrichment analysis were used to assess putative biological processes (BP), molecular functions (MF), and cellular components (CC) associated with genes of interest. KEGG analysis defined the pathways associated with SCAMP2/4/5 functions and their co-expression genes. Analyses utilized the human genome as a background parameter. P < 0.05 was the significance threshold.The DAVID v6.8 (http://metascape.org) was employed for process and pathway enrichment analyses of genes co-expressing with SCAMP2/4/5 in leukemia. As an effective, efficient and user-friendly gene-list analysis tool, Metascape enables multi-platform analyses of multi-omic data28. Only terms meeting the following criteria were deemed significant: p < 0.01, minimum count 3, and enrichment factor > 1.5. The core protein–protein interaction (PPI) network was constructed by BioGRID, InWeb_IM and OmniPath, and the results were visualized with Metascape. Then MCODE  cluster analysis was performed to detect key MCODE clusters via setting parameters to the most central and densely connected clusters in the PPI network, those being degree cutoff = 2, node score cutoff = 0.2, K-score = 2 and MAX depth = 100. Further, the functions of the most significant modules chosen from the PPI network were predicted using Metascape at a significance of p < 0.05.Metascape  and mutation (missense mutation and truncating mutation) , with Pearson’s correction involved. The results demonstrated significant and positive correlations between SCAMP genes expression Fig. C.29, acute adult T-cell leukemia (ATL) datasets from Choi’s dataset30, and AML datasets, T-cell Acute Lymphoblastic Leukemia (T-ALL) datasets and B-cell Acute Lymphoblastic Leukemia (B-ALL) datasets from Andersson’s dataset31. SCAMP1 was overexpressed in childhood ALL (fold change (FC) = 1.727) from Coustan-Smith’s dataset, and in acute ATL (FC = 1.505) relative to normal controls from Choi’s dataset. The expression of SCAMP1 was significantly higher in AML (FC = 1.786), T-ALL (FC = 1.537), and B-ALL patients (FC = 1.583) from Andersson’s dataset  transport to cell surface and cooperate to prevent HER2 + /ER-/PR- breast cancer invasion, indicating SCAMP1 as a tumor repressor in breast cancer4. However, in the GEPIA database, SCAMP1 also showed a trend of increased expression in AML patients compared to normal controls, but without significant difference. Thus, the diagnostic role of SCAMP1 needs to be further clarified.We found that SCAMP1 expression levels showed increased in AML patients compared to normal controls in Andersson Datasets, which is consistent with a previous report34. Overexpression of SCAMP3 was found to be closely related to poor overall survival in human hepatocellular carcinoma and that knockdown of SCAMP3 decreased cell proliferation and cell cycle progression of HCC cells11. This suggested that SCAMP3 may play a carcinogenic role. However, SCAMP3 was also reported to function as a novel tumor suppressor in lung cancer by modulating EGFR signaling and cytokinesis12. Herein, the expression of SCAMP3 in AML tissues was found higher than normal controls from Oncomine database. Despite there being no correlation between expression of SCAMP3 and prognosis of AML in our study, SCAMP3 may play an important role in regulating AML.SCAMP3, an important membrane carrier, was reported to participate in cell growth by interacting with DRAM-1, which was in turn involved in the activation of mTORC135. However, SASP was a major trait of senescent cells36. In addition, an in vitro study demonstrated that senescent fibroblasts overexpressing SASP may potentially stimulate or accelerate neoplastic progression by creating a tumorigenic microenvironment37. Similarly, the growth of leukemia cells created abnormal bone marrow microenvironments, which played a key role in the initiation and development of hematological malignancies38. This raised the question if SASP plays the same role in leukemia. In our report, we found that a higher SCAMP4 expression significantly correlated with poor overall survival, pointing to SCAMP4 as a possible tumor promoter. The molecular mechanism involved in SCAMP4 in AML and whether elevated SCAMP4 in AML promotes the secretion of SASP is the next step that needs to be investigated.Previous research has reported that overexpression of SCAMP4 promoted the secretion of senescence-associated secretory phenotype (SASP) factors and also affected cell proliferation of WI-38 cells expressing SCAMP4-Myc, in contrast to cells only expressing the Myc tag41. Similarly, SCAMP5 is directly involved in calcium-regulated exocytosis of signal peptide-containing cytokines via co-distributing and compelling with SNAREs10. Interestingly, increasing evidence has proven that SNAREs played a core role in vesicle transport by vesicle-target membrane fusion, which was vital for compartment integrity, exocytosis and trafficking within the cell47. In addition, SNAREs play an essential role in the delivery of mutant KRAS from recycling endosome to plasma membrane through vesicular transport, which facilitated KRAS associating with downstream effectors to carry out its tumorigenic action47. Nevertheless, whether the docking sites of SNARE and SCAMP2/5 are involved in regulating other important signal transmission remained obscure. Herein, SCAMP2 and SCAMP5 were found significantly overexpressed in AML, and their expression showed positive correlation with each other. Importantly, high SCAMP2 and SCAMP5 expression was significantly associated with poor overall survival in AML patients, indicating the oncogenic roles of these transcriptional factors in AML. Further experimentation involving the docking sites of SNAREs and SCAMP2/5 may uncover the molecular mechanism of AML.Until now, little was known about the expressions and specific roles of SCAMP2 and SCAMP5 in AML. SCAMP2 was shown to set up soluble N-ethylmaleimide sensitive factor attachment protein receptors (SNAREs) interactions and has an essential function in granule exocytosis by fusion pore formation50. In AML, the activation of this pathway, can bring adversely prognostic impact to AML patients53.Taken together, these data suggest SCAMP2/4/5 may be potential prognostic biomarkers for AML.We constructed a network of SCAMP2/4/5 and 50 closest co-expressed genes for each of them. The results of the functional analysis of distinct SCAMP and its co-expressed genes indicated that these genes were involved in multiple pathways related to tumorigenesis and progression, such as human T-cell leukemia virus 1 (HTLV-1) infection, acute myeloid leukemia, and mTOR signaling pathways. The mTOR, downstream effector of PI3k, can make leukaemia-initiating cells acquire the properties of proliferation and survival and eliminate haematopoietic stem cell54. Our data including the potential mechanism involving NF-κB pathway suggest a vital role of SCAMP2/4/5 in tumorigenesis and progression of AML.We also performed functional enrichment analysis of overlaps of SCAMP2/4/5 and their co-expressed genes in Metascape to further prove the results of functional enrichment analysis above and explore the potential interaction mechanisms of SCAMP2/4/5 in AML. One of the interesting gene set was involved in NF-kappa B signaling pathway. In addition, NF-κB pathway, which was proved to be strongly correlated with TNF-α signaling, accounts for the progression of AMLIn summary, our results suggest that SCAMP2/4/5 are potential prognostic markers for AML, and that SCAMP2 and SCAMP5 individually or in combination may be used as diagnostic markers for AML.


This paper provides a comprehensive review on the use of infrared thermography to detect delamination on infrastructures and buildings. Approximately 200 pieces of relevant literature were evaluated, and their findings were summarized. The factors affecting the accuracy and detectability of infrared thermography were consolidated and discussed. Necessary measures to effectively capture latent defects at the early stage of delamination before crack formation were investigated. The results of this study could be used as the benchmarks for setting standardized testing criteria as well as for comparison of results for future works on the use of infrared thermography for detection of delamination on infrastructures and buildings. With the aging of civil infrastructures and buildings, those deterioration has become an important social issue that can threaten public safety. The American Road & Transportation Builders Association reported in 2020 that 36% of bridges in the US need replacement or rehabilitation due to their aging . SimilarDefects in infrastructures can be diverse and include delamination, cracks, staining, and spalling, caused mainly by water penetration, reinforcement corrosion, thermal/moisture movements, differential settlement/loading, poor construction practices, etc. ,8. AmongIn recent decades, various NDTs were developed to detect defects in multiple fields since they can evaluate object characteristics ,16,17. EIRT is defined as a process of measuring surface temperature distribution using IR cameras and processing and interpreting the data of IR images . For infThis paper focuses on the use of IRT to detect delamination on infrastructures and buildings to prevent falling objects from heights that endanger public safety. It provides a comprehensive review of the use of IRT by providing backgrounds, principles, and state-of-the-art knowledge on affecting factors and desirable conditions. This paper will contribute to increasing the reliability of IRT and facilitating further research.This section investigates review papers on IRT within the last decade. Recent review papers on IRT were conducted from perspectives of applications, methodologies, and research trends.The first perspective is IRT applications, which are commonly used in reviews. Application reviews range from the level of introducing case studies in industrial fields to the level of in-depth investigation of a specific application. IRT applications were developed in many fields, including medical , aerospaSeveral review papers focused on the energy audit of building envelopes using IRT to evaluate building energy performances ,48,50,70The second perspective is the methodologies of conducting IRT. A wide variety of methodologies was developed and is classified based on their features and principles . One claThe final perspective is the analysis of research trends based on statistical data of the number of past studies. For example, Fox et al.  analyzedAs mentioned above, previous reviews about IRT were conducted from various perspectives. However, no review focuses on the characteristics of detectable delamination and measurement conditions affecting reliabilities and detectability of IRT for infrastructures and buildings.bW is the total radiant flux emitted per unit area (W/m2), σ is the Stephan–Boltzmann constant, and objT is the absolute temperature of the object (K). The black body is defined as an ideal object that absorbs all the radiation that collides with it at any wavelength. However, an actual object, called a grey body, is not a black body because objects have some reflection and transmission. The radiant energy emitted by a grey body (gW) is as follows:ε is emissivity, defined as the ratio of the radiant energy emitted from the object to the energy emitted from the black body at the same temperature.This section explains the principle of temperature measurement by IR cameras. Heat energy can be transferred in three ways: conduction, convection, and radiation. Temperature measurement by IR cameras utilizes radiation transfer. All objects with an absolute temperature greater than 0 K emit electromagnetic waves, mainly in infrared spectra. According to Stefan–Boltzmann’s law, radiant energy from a black body is as follows:objW), assuming the object as the black body, emission from surroundings reflected on the object (reflW), and emission from the atmosphere (atmW). The following formula expresses the total heat radiation detected by the IR camera :τ is the transmittance of the atmosphere. The reflected radiation assumes that reflection temperature reflT is the same for all reflections from surroundings, and the emissivity of surrounding surfaces reflε is assumed to be one. Radiation from the object and reflected radiation are absorbed by the atmosphere during traveling. Atmospheric radiation is emission from the atmosphere between the object and the camera at ambient temperature atmT. “1 − τ” indicates the atmosphere’s emissivity, and τ depends on atmT, relative humidity, and measurement distance between the target object and the IR camera. Therefore, accurate surface temperature measurements need to be compensated for the effects of emissivity ε, ambient temperature atmT, relative humidity, reflection temperature reflT, and distance [IR cameras capture radiant energy in a specific infrared spectrum region emitted from an object and convert the energy into a temperature value. distance .In particular, the emissivity of target objects has a significant influence on temperature measurement . The emiThere are various methodologies of conducting IRT. They are classified from several viewpoints: analysis scheme, mode, and measurement method . When asAn analysis scheme is a classification criterion based on the origin of the energy input to a target object to generate temperature distribution on the surface. There are passive and active IRT.Passive IRT utilizes natural heat sources as stimuli to generate a thermal gradient inside an object, causing thermal contrast on the object’s surface between sound and defect area . The priActive IRT uses artificial heat sources to heat a target object to generate a thermal gradient . Active Active IRT is further classified according to heating processes: pulsed IRT (PT), step heating thermography (SH), and lock-in IRT (LT) . PT provIRT is also classified into two modes according to the relative position of an IR camera and a heat source: transmission mode and reflection mode ,66. ThesThe transmission mode places a heat source on one side and an IR camera on the opposite side of a target object . The temThe reflection mode places a heat source and an IR camera on the same side of a target object . In thisIRT has two measurement methods: qualitative and quantitative IRT . QualitaQuantitative IRT is a numerical evaluation method by comparing temperature values on IR image pixels between identical items or baselines . The quaIRT for detecting subsurface delamination on infrastructures and buildings generally adopts passive IRT of the reflection mode . The priFrom early morning to noon, solar radiation increases, and the ambient temperature rises. Solar irradiation and warm ambient temperature heat the surface of a target object, creating heat flow to the inside of the object. The thermal conductivity of concrete is approximately 1.6 W/mK, while that of air, filling delamination, is significantly low at 0.024 W/mK . Thus, dOn the other hand, during nighttime, the surface temperature declines due to radiative cooling and low ambient temperature . The heaIRT survey can be conducted during both the heating cycle and the cooling cycle. However, the periods when the two cycles exchange in the early morning and the evening, called interchange times , are notAnalysis methods for delamination detection from IR images were developed as postprocessing procedures . The anaOne-time data analysis, called single-frame image processing , process(a)Visual EvaluationVisual evaluation is a method that an inspector interprets temperature distribution patterns in an IR image by comparing surroundings and assesses the presence and location of delamination ,96,97. T(b)Thermal ContrastThermal contrast, called ΔT, is referred to the surface temperature difference between the delamination area and the sound area ,100. The(c)Image ProcessingImage processing was developed to extract temperature abnormalities automatically, quantitatively, accurately, and sensitively. The processing mainly utilizes threshold temperature values and temperature gradients.Threshold temperature values are generally used for image processing. This image processing sets a threshold temperature value to judge areas as delamination and converts an IR image into a binary image based on the value. The processing has the advantages of simple evaluation; however, this processing primarily has two challenges.The first challenge is determining threshold values because the values are affected by environmental conditions. Therefore, various methods to decide threshold values were proposed. The primitive method is that an inspector decides a threshold value that gives clear contrasts between sound and delamination areas by changing the value. The disadvantages of this method are subjective and time-consuming . JapanesThe second challenge of threshold values is difficult to evaluate the entire target object by one global threshold value. The reason may be that the entire surfaces of infrastructures or buildings are not under the same conditions, and each local area has a different average temperature and gradient . Thus, mTemperature gradients are also employed for image processing. The processing identifies the areas of thermal anomalies based on the significant temperature changes at the edge of delamination. The advantages of the gradient are that measuring accurate temperature values is not required , and a sOverall, A substantial number of image processing methods using one IR image were developed. However, they may be designed to be optimized under specific conditions. To improve the accuracy of detectability and applicability for field inspection, further research is needed.Time-series data analysis collects courteous IR images over time and analyses time-series temperature data. It is also called time-lapsed thermography ,75, timeRegarding environmental conditions, four factors are generally stipulated: solar irradiation, ambient temperature, wind, and weather. All documents recommend the survey with direct sunlight exposure because solar irradiation has significant energy input and generates high thermal contrast. ASTM  for bridIR camera specifications, distance from a target object to an IR camera, and observation angle are also mentioned in the documents. One of the specifications is temperature resolution, represented as noise equivalent temperature difference (NETD) . The NETIn addition to IRT, several NDTs were developed to detect delamination on infrastructures and buildings: audio methods, stress wave methods, and electromagnetic methods ,122,123.Audio methods are based on a feature that when a mechanical impact is applied on a target object from outside, delamination areas produce impact sound with a frequency significantly different from intact areas (hollow sound); an inspector listens to the impact sound and evaluates delamination areas. The suitable method of giving mechanical impact relies on target objects. For vertical surfaces such as buildings or tunnel linings, coin tapping testing, which uses coins, steel rods, or lightweight hammers, is widespread ,122. ForStress wave methods utilize characteristics of stress-wave propagation in a target object. Among the methods, impact-echo testing (IE) and ultrasonic testing (UT) can detect delamination. IE is a method that a mechanical impact is applied to a target object, then the frequency of the wave reflected on delamination is analyzed with FFT ,127. UT Ground penetration radar (GPR) employs electromagnetic pulses . ElectroIRT was compared with these NDTs about delamination detection: coin tapping testing , chain dThe detectability of IRT is affected by many factors, including environmental conditions, delamination properties, target objects, and IR cameras . Thus, tTest methods used in the previous studies are classified into four categories: laboratory test, outdoor test, field survey, and numerical simulation. In one laboratory test, specimens are prepared with polystyrene foam plates or other Insulation materials embedded to imitate delamination. An outdoor test places specimens with simulated delamination in an outdoor location and observes the specimens for several days . ThermalA field survey is a method of inspecting existing infrastructures or buildings. The survey is often used to verify the results of laboratory tests and outdoor tests ,148. TheNumerical simulation or modeling may provide useful information on the impact of factors such as irradiation ,170, defThe results of outdoor tests and field surveys may rely on the test region and surface direction. Most research was conducted under temperate climates, with high daily temperature changes and stable weather, for example, in the US  and EuroSurface direction is also an important test condition because it relates to the magnitude and time of solar irradiation. In outdoor tests and field surveys, surface directions were mainly horizon or south elevation in the Northern Hemisphere. The horizontal direction assumed bridge decks, and the south elevation is considered ideal conditions with solar irradiation in buildings.Although metrics and criteria of detectable delamination are critical to identifying delamination and evaluating the impact of factors, there are no unified metrics and criteria. The metrics commonly used in previous studies are thermal contrast and signal-to-noise ratio (SNR).Thermal contrast or ΔT, the temperature difference between sound and delamination area, is the most commonly used matric because it is simple and easy to analyze. However, the criterion of ΔT to be judged as delamination significantly differ depending on the studies, ranging from 0.2 to 1.2 °C. For example, Hiasa et al. ,159 and Meanwhile, some studies proposed multiple criteria of ΔT depending on test methods. For example, Zheng et al.  mentioneSNR, which is used in engineering, compares single levels of a target area to signal levels of background noise, calculated by the following equation [areaS is the average temperature value in the delamination area, areaN is the average value in the surrounding area, and noiseσ is the standard deviation in the surrounding area. Positive SNR means detectable delamination, and negative SNR means undetectable. The advantage of the SNR is that because of signal level evaluation, the metric and criterion can be applied not only to raw IR images but also processed images, such as PPT or PCT [The SNR is utilized as the metric to evaluate the detectability of delamination ,85,161 oT or PCT .This section compares and synthesizes previous studies on factors affecting delamination detectability. Suitable time windows to conduct passive IRT are critical information for getting proper IR images to analyze. Multiple environmental factors, such as irradiation and ambient temperature change, can affect thermal contrast intricately. Thus, time windows are generally examined by outdoor tests and numerical simulations. Regarding horizontal surface and south elevation, available time windows proposed by the literature are generally around 10 a.m. to 3 p.m. due to the presence of solar irradiation ,132,147.Furthermore, several studies proposed that suitable time windows relied on delamination depth. The reason is that the deeper delamination is, the longer it takes for heat flow to reach delamination. Washer et al.  showed tMeanwhile, several studies have focused on interchange times, which can not detect delamination due to low thermal contrast. Edis et al.  calculatOverall, many studies examined suitable time windows for horizontal surface and south elevation during daytime. Although it is affected by delamination properties, the suitable time window is generally noon to early afternoon when delamination depth is under 6 cm.Regarding east and west elevation, time windows with solar irradiation on the elevation should be optimal. It means that the suitable time for east elevation is in the morning and that for the west elevation is in the afternoon. For example, Buare et al.  observedNighttime or the cooling period is a candidate for suitable time windows; however, this is still being debated. One opinion is that nighttime is not appropriate or impossible to conduct IRT. Yehia et al.  failed tFurthermore, there are still two opinions about suitable time windows during nighttime: early night or early morning. Hiasa et al.  mentioneShaded areas, soffit, or north elevation, which has no solar irradiation on the inspected surface, may exist on infrastructures and buildings. In these areas, suitable time windows during daytime are generally around noon due to the peak of ambient temperature; however, these time windows are shorter than those of sunny areas ,103,158.As explained above, suitable time windows for IRT proposed by previous studies are not consistent. The reason is that the windows are affected not only by surface direction but also by environmental conditions and delamination properties. Therefore, investigating suitable time windows for each region and the target object is required to conduct IRT properly.2 on a south elevation and 1300 W/m2 on a horizontal surface at noon [Solar irradiation is a primary stimulus producing heat flow ,145. It  at noon ,148,159. at noon ,169,170. at noon ,148,162. at noon ,167. Mea at noon ,91. Over2 was required for 5.1 cm deep delamination to generate the detectable thermal contrast of 1 °C based on statistical analysis. Likewise, Raja et al. [2 produced the thermal contrast of 1 °C for 6.3 cm deep delamination based on the numerical simulations. In addition, the authors stated that a heat flux rate greatly influenced thermal contrast, especially for shallow and small delamination. These studies indicate that total irradiation of approximately 700 Wh/m2 could be required to conduct passive IRT.Few studies quantitatively investigated the relationship between irradiation and thermal contrast. Washer et al.  conductea et al.  proposedDaily ambient temperature change is one of the drivers to generate thermal contrast due to convection heat transfer. The daily change is a primary heat source in shaded areas or under cloudy weather . MultiplAmbient temperature values might influence thermal contrast. Tran et al.  mentioneFor buildings, an ambient temperature difference between indoor and outdoor can also affect thermal contrast. Edis et al.  conducteWind velocity is an environmental factor to be considered when performing passive IRT, as it relates to convection heat transfer . High wiUnder the condition of solar irradiation or during the heating cycle, high wind velocity decreases thermal contrast. The reason is that the surface temperature of a target object is generally higher than ambient temperature, so that the wind removes heat energy from the surface. For example, Washer et al.  statistiIn contrast, in shaded areas, high wind velocity could increase thermal contrast. The reason is that the surface temperature is generally lower than ambient temperature, and high wind velocity increases energy input from the air to the object’s surface. Washer et al.  pointed Relative humidity (RH) is considered to affect thermal contrast due to two theories. One theory is that high RH increases convection heat transfer between the object surface and atmosphere . Thus, iHowever, the effect of RH may be limited and not significant. For example, Tran et al.  argued tWeather is closely related to other environmental factors. A sunny day is optimal for IRT regardless of sunny or shaded areas due to high solar radiation and high daily ambient temperature change ,147,162.A method to predict thermal contrast from meteorological data was proposed. Watase et al.  proposedSeasons are related to the amount of solar irradiation and daily ambient temperature change. Hence, the effects of seasons were examined using numerical simulation, but the results are not consistent. Hiasa et al.  concludeDetecting small delamination at the early stage of deterioration leads to ensuring public safety. As shown in Delamination size is also related to the response time of the maximum thermal contrast. Maierhofer et al.  confirmeDepth from delamination to the surface significantly affects thermal contrast. Thus, detectable depth is an essential indicator of IRT abilities. Approximately 78% of the studies include depth as study parameters, as shown in It is not easy to detect deep delamination as deep delamination has low thermal contrast. Furthermore, detectable depth was a controversial and much-disputed subject even under the same test condition. In (b) and (c) outdoor tests with solar irradiation, Washer et al.  mentioneDepth estimation was also of great interest to researchers because depth is essential information to evaluate severity. For example, AASHTO Guide Manual for Bridge Element Inspection  assessesThe estimation method based on response time utilizes that delamination depth correlates with the time from energy input to the generation of thermal distribution on the surface . In laboThe estimation method based on thermal contrast magnitude uses the correlation between thermal contrast and depth. Tran et al.  showed iAs described above, depth estimation methods using response time or thermal contrast magnitude are possible under constant or controlled conditions such as laboratory tests. However, since environmental conditions fluctuate, further research is needed to estimate depth under outdoor conditions.Delamination width and depth are closely related to detectability while interacting. It is generally considered that the minimum detectable width is at least 1–2 times the depth or more . Thus, mMore specifically, each condition has a different tendency for detectable delamination distribution and WTDR criteria. In (a) laboratory tests, the distribution results are almost consistent among the literature compared to outdoor tests. This is probably because such laboratory tests can optimize energy input and remove unintended noise from the surroundings. Additionally, the detectable and undetectable regions are relatively separated by a straight line. Thus, the WTDR criteria proposed by the literature are relatively low values of 1.11–1.43 ,85. ThisAs described above, the WTDR criteria of detectable delamination are influenced by test methods, the presence of solar irradiation, measurement cycle, and test regions. As a result of integrating previous studies, WTDR criteria are approximately 1.25 in (a) laboratory test, 2.0 in (b) outdoor test with irradiation during the heating cycle, and 2.5 in (c) outdoor test with irradiation during the cooling cycle and (d) outdoor test in shaded areas.Delamination thickness is also a factor to consider for its impact on detectability. Previous studies have generally set the thickness of 0.1–2 cm by adjusting the thickness of embedded materials. Thick delamination has a low overall heat transfer coefficient; thus, it generates significant thermal contrast regardless of environmental conditions or measurement cycles ,146,159.Meanwhile, thickness is considered the minor effect on thermal contrast among the geometric factors of delamination . Hiasa eDelamination is usually filled with air; thus, the thermal properties of delamination are considered to resemble the air. However, making air-filled delamination in a concrete specimen with a predetermined size and depth is not easy, except for delamination beneath tiles. To simulate delamination, materials with low thermal conductivity are embedded in specimens. Thus, several studies have examined the effect of embedded materials ,141,168.The materials of the target object affect thermal contrast because heat flow is determined by thermophysical properties of the materials: thermal conductivity, specific heat capacity, and density. The properties of concrete change depending on compression strength and mix proportions. For example, Rocha et al.  and FarrBuilding walls are generally layered with different materials rather than the single material of concrete. When finish materials are the same, substrate materials can also affect thermal contrast. Lourenço et al.  examinedOther factors related to target objects investigated by the previous studies include rebars, water penetration, and surface conditions. The effects of these factors might be inevitable when inspecting existing infrastructures and buildings.Rebars are usually embedded in concrete parallel to the surface to reinforce concrete structures. Rebars have a high thermal conductivity of 12.5 W/mK, much higher than concrete of 1.8 W/mK. Therefore, rebars may diffuse heat flow parallel to the surface, resulting in low thermal contrast. According to laboratory tests and outdoor tests, the effect of rebars is different depending on the relative position of rebars and delamination ,147,167.Water penetration or high moisture content in a target object generates nonuniform temperature distribution on a surface due to three physical phenomena: evaporative cooling ,162,177,Surface conditions, such as color and obstacles on the surface, affect IR images. Building facades are generally colored with paint or colorful materials. Lourenço et al.  studied The surfaces of infrastructures and buildings are not always clean and may have small obstacles. Hiasa et al.  stated tTwo types of IR cameras are generally used for IRT: a short-wavelength (SW) camera and a long-wavelength (LW) camera . Table 5SW cameras use a cooled quantum detector sensitive to high-energy emissions from hot objects . Thus, tLW cameras use an uncooled microbolometer detector sensitive to low-energy emissions. Thus, the quality of IR images is relatively high when a target is at a low temperature. In addition, LW cameras are less subject to solar reflections on surfaces. In contrast, the cameras are susceptible to reflections of surrounding buildings and the sky on glazed tiles or smooth surfaces ,97,160. Regarding the influence of IR camera type and model, Hiasa et al. ,159,160 A short distance from the IR camera to the target object is considered ideal ,160,162;Atmospheric attenuation is a phenomenon in which water vapor and carbon dioxide in the atmosphere absorb IR . Due to The size of a captured area may influence the efficiency of IRT surveys and detectability. The area captured is determined by an IR camera’s field of view (FOV) and distance. FOV indicates the largest area that an IR camera can capture, described in horizontal and vertical degrees, and is determined by the focal length and the detector size of the IR camera. A long-distance measurement can capture a large area at once and improve efficiency. However, this IR image tends to include surroundings or nontarget objects with high or low temperatures. IR cameras automatically adjust the span of the temperature color scale based on the maximum and minimum temperature in an IR image. Therefore, Lourenço et al.  stated tTo measure the temperature value of a small area accurately, at least a smaller spatial resolution than the area is required . SpatialAs described above, distance is related to detectability from the aspects of atmospheric attenuation, captured area, and spatial resolution. It is desirable to capture IR images as close as possible while balancing productivity and limitation of accessibility.An observation angle could affect temperature values measured by IR cameras and detectability. This is because the emissivity of objects relies on the angle with respect to the surface. In general, the emissivity of nonmetallic materials is stable from the angles of 0° to 45° and decreases at higher angles ,194. SevAt angles above 45°, the detectability of IRT may decline because of thermal contrast reduction or reflection noise. Scott et al.  reportedWhen surveying a wide area, mounting an IR camera on a platform can enhance the IRT survey’s efficiency compared to by hand. For example, in the bridge deck inspection, an IR camera fixed to the top of a car continuously captures a road lane . HoweverRecent developments in robotics allow inspectors to use unmanned aerial vehicles (UAVs) as a platform to inspect infrastructures and buildings ,196,197.Capturing latent defects at the early stage of delamination even before delaminated objects falling is essential for integral components of infrastructures and buildings. With this in mind, a comprehensive review on the use of IRT to detect delamination on infrastructures and buildings was presented.Three classifications of IRT for assessing defects were explained to clarify the methodologies used in delamination detection. Regarding delamination detection, the principle, evaluation protocols with one-time and time-series data analysis, and standards and guidelines were consolidated. Additionally, the performance of IRT in detecting delamination was compared with that of other NDTs.Suitable time windows for the inspection depend on the direction of the inspection surface and delamination depth. For shallow delamination on a horizontal surface or south elevation, the windows are noon to early afternoon and late evening to early night.A large amount of total solar irradiation is desirable because irradiation is the primary heat source to generate thermal contrast.High daily ambient temperature change allows IRT even in shaded areas.A low wind velocity is preferable in sunny areas.Fine weather is optimum for the heating and cooling cycles because of solar irradiation, high daily ambient temperature changes, and radiative cooling.Delamination of large size has high thermal contrast and is easy to detect.The detectable depth of delamination is greatly affected by environmental conditions. Delamination of at least 3–5 cm or less could be detected in outdoor conditions.The width to depth ratio (WDTR) of delamination also affects detectability. The WTDR criteria of detectable delamination are 1.25 under laboratory conditions and 2–2.5 under outdoor conditions.The target object with high thermal conductivity has high thermal contrast, and the detectability is low on the insulation walls or low-strength concrete.Water penetration into delamination causes the opposite behavior of the thermal contrast of delamination.Dark color surfaces in sunny areas are advantageous for inspection.The influence of obstacles on the surface can be removed by complementing IR images with visual images.Both types of SW cameras and LW cameras can be used for inspection. An appropriate type should be selected according to the noise of the surrounding environment.The close distance from an IR camera to a target object is desirable in terms of atmospheric attenuation, captured area, and spatial resolution while balancing productivity and limitation of accessibility.When IR camera platforms, such as vehicles or UAVs, move quickly, SW cameras can collect clear IR images compared with LW cameras.Experimental methodologies employed by studies over the last 20 years on factors affecting delamination detection were discussed. Furthermore, the impact of factors on detectability was also investigated. Factors studied include environmental conditions, delamination properties, target objects, and IR cameras. Although the results of the studies were not always consistent due to the differences in experimental conditions, general desirable conditions for IRT are summarized below:The results of this study could be used as the benchmarks for setting standardized testing criteria, as well as for comparison of results for future works on the use of infrared thermography for detection of delamination on infrastructures and buildings.


P = 2.15 × 10−8; KMT5B). We also identify a genome-wide significant risk locus for the cytogenetically normal AML sub-group (N = 1287) at 6p21.32 . Our results inform on AML etiology and identify putative functional genes operating in histone methylation (KMT5B) and immune function (HLA).Acute myeloid leukemia (AML) is a hematological malignancy with an undefined heritable risk. Here we perform a meta-analysis of three genome-wide association studies, with replication in a fourth study, incorporating a total of 4018 AML cases and 10488 controls. We identify a genome-wide significant risk locus for AML at 11q13.2 (rs4930561;  Genome wide association studies in cancer are used to understand the heritable genetic contribution to disease risk. Here, the authors perform a genome wide association study in European patients with acute myeloid leukemia and identify loci associated with risk of developing the disease. Rare germline variants in transcription factors and other genes regulating hematopoietic cell differentiation and proliferation are highly penetrant for AML3, with some causative for debilitating human syndromes that include AML as a component, while others cause familial AML with high penetrance. Although such variants demonstrate a role for genetics in AML susceptibility, they are rare and do not make a major contribution to population disease burden4. A modest increased risk of myeloid malignancy for first-degree relatives of non-familial AML patients further supports a role for genetics in disease susceptibility5, which for the majority of cases is likely determined by co-inheritance of common low penetrance variants consistent with the multigenic model for complex human diseases7.Acute myeloid leukemia (AML) is the most common acute leukemia in adults of European ancestryOutside of highly penetrant syndromic or familial disease, genetic susceptibility to AML remains largely unexplained. To identify AML risk loci we conducted three genome-wide association studies with participants of European ancestry, with replication in a fourth study. Here we report the identification of risk loci for AML, including pan-AML irrespective of disease sub-type and for cytogenetically normal AML. These data inform on disease etiology and demonstrate the existence of common, low-penetrance susceptibility alleles for AML with heterogeneity in risk across sub-types.http://www.haplotype-reference-consortium.org/)8 was used to derive estimated genotypes for >7 million single nucleotide polymorphisms (SNPs) for each study. We combined the association test statistic for 6694056 autosomal SNPs common to all three GWAS after exclusion of those with an INFO  score of <0.6 and a minor allele frequency (MAF) of <0.01, and conducted a meta-analysis under a fixed-effect model for variants associating with AML. Given the genetic and biological heterogeneity of AML, sub-type specific odds ratios (ORs) were also calculated for the largest AML sub-group, cytogenetically normal AML (N = 822), using data for 6532175 SNPs common to all three studies. Quantile-quantile plots of observed versus expected P values  >0.01) for all AML cases and cytogenetically normal AML cases showed minimal inflation of test statistics across all three GWAS after adjustment for nominally significant principal components in each GWAS   and 4q22.3 (rs75391980) that surpassed genome-wide significance (P ≤ 5 × 10−8), and additional suggestive associations (P < 10−6) at 2q36.1 (rs4674579), 3q (rs2621279), 5q35.1 (rs13183143), 11q13.2 (rs11481), and 20p12.3 (rs6077414) for all AML and at 9p21.1 (rs10969985) and 13q21.31 (rs143280565) for cytogenetically normal AML . Following implementation of rigorous quality metrics to each GWAS  showed minimal inflation of test statistics   two of the associations identified in the discovery GWAS meta-analysis, with consistent direction and magnitude of effect sizes across all four studies, including 11q13.2  for all AML irrespective of sub-type (N = 4018) and 6p21.32  for cytogenetically normal AML (N = 1287). Meta-analysis of SNPs common to all four GWAS  also revealed additional borderline significant susceptibility loci at 1p31.1  for all AML and at 7q33  for cytogenetically normal AML, both with consistent direction and magnitude of effect across all four studies . Following the application of SNP and sample quality control metrics  for association with AML for any of the risk variants across the 4 GWAS studies  within 500 kb of the lead variant I2 = 78.47, PQ = 0.03)  (Supplementary Table N = 767) or those with cytogenetically normal AML (N = 369)  from the UK, Germany, and Hungary. However, none of the 4 AML susceptibility variants identified here were significantly associated with either relapse-free or overall survival in univariate analysis that included all AML patients (BICRA gene (rs75797233)10 was not significantly associated with AML risk in our study  1.06, 95% CI 0.87–1.29). However, this variant was imputed to sufficient quality (INFO > 0.6) in only three GWAS studies and statistical power to detect an association with AML for this relatively uncommon variant (MAF 0.02) is compromised.A previously reported susceptibility variant for AML in the P = 2.15 × 10−8) which maps to the KMT5B gene on 11q13.2 , RP5-901A4.1 (PBH = 4.1 × 10−24), ALDH3B1 (PBH = 4.48 × 10−18), IGHMBP2 (PBH = 2.85 × 10−16), RP11-802E16.3 (PBH = 6.76 × 10−15), and CHKA (PBH = 1.73 × 10−14), DOC2GP (PBH = 1.32 × 10−12), TCIRG1 (PBH = 7.83 × 10−5), NDUFS8 (PBH = 1.69 × 10−4), UNC93B1 (PBH = 1.09 × 10−3), PP1CA (PBH = 0.01), and RP11-554A11.9 (PBH = 0.01). rs4930561 is not, however, an eQTL for KMT5B (PBH = 0.99) , which maps to the HLA locus on 6p21.32, intergenic to HLA-DQB1 and HLA-DQA2 within the major histocompatibility (MHC) class II region . HLA-DQB1*03:02 (r2 = 0.56 with rs3916765) was significantly under-represented in AML cases compared to controls in meta-analysis , identifying this as the most likely classical HLA allele underpinning the association with AML at the 6p21.32 locus  was also significantly under-represented in cases compared to controls  , consistent with linkage disequilibrium between these two alleles (r2 = 0.50).We identified strong statistical evidence for an association with cytogenetically normal AML for rs3916765  and FLT3-mutated AML  compared to NPM1-wildtype AML  or FLT3-wildtpe AML   which maps to a block of linkage disequilibrium upstream of the CACHD1 gene on chromosome 1p31.3  and AK4 (PBH = 1.26 × 10−2) where the AML risk variant is associated with higher expression of AK4 and lower expression of RAVER2 , which maps close to the AKR1B1 gene on chromosome 7q33   encodes a lysine methyltransferase that is frequently mutated in human cancers, with gene amplifications being particularly common15. KMT5B is implicated in AML pathogenesis where mutation has been associated with transformation from precursor myelodysplastic syndrome to AML16. Mutations in other lysine methyltransferases such as KMT2A (MLL1) occur with high frequency in AML17. Although KMT5B is a strong candidate for an AML susceptibility gene a priori we cannot exclude mechanisms involving other local genes. For example, the AML risk variant at the 11q13.2 susceptibility locus is significantly associated with lower expression of CHKA, which encodes a protein involved in phosphatidylcholine biosynthesis. CHKA is significantly upregulated in mouse haematopoietic stem cells and human leukemia cell lines upon restoration of TET2 function18, a tumor suppressor which blocks aberrant self-renewal and which is frequently mutated in AML resulting in loss of function19.By conducting a meta-analysis of three large genome-wide studies with validation in a fourth study, we identify four susceptibility loci for AML, demonstrating the existence of common, low-penetrance susceptibility alleles for this genetically complex disease. Specifically, our data identify a major susceptibility locus for AML at the 11q13.2 cis-eQTL for RAVER2 and AK4. RAVER2 encodes a ribonucleoprotein involved in RNA splicing where expression associates with transformation from myelodysplastic syndrome to AML20. In both human and mouse hematopoietic stem cells RAVER2 is identified as a target gene for miR-9921, which regulates normal and malignant hematopoietic stem cell self-renewal22 and expression is significantly associated with prognosis in AML22. AK4 belongs to the adenylate kinase (AK) family of proteins that catalyze the phosphorylation of nucleotide monophosphate precursors to their di- and triphosphate forms. AK4 is localized to the mitochondria and is a target gene for hypoxia-inducible factor 1 alpha (HIF-1α)23, an established tumor suppressor in human and murine AML25.We also report a putative pan-AML susceptibility locus at 1p31.3 that is HLA gene. This region carries susceptibility alleles for numerous human cancers including hematological malignancies such as chronic lymphocytic leukemia27 and Hodgkin lymphoma28, where risk is mediated via differential antigenic presentation/T-cell receptor recognition or altered risk of oncogenic infection. Somatic dysregulation of antigenic presentation through altered gene expression, loss of heterozygosity or genomic deletion is a common feature of many human solid cancers leading to failed immune surveillance31. Somatic loss of HLA alleles is rare in AML at disease presentation although it has been reported as a mechanism of immune escape after bone marrow transplant leading to relapse35. Our data identify the HLA-DQB1*03:02 and HLA-DQA1*03:01 alleles as significantly under-represented in cytogenetically normal AML cases compared to controls. Cytogenetically normal AML is characterized by somatic mutation in genes such as NPM1 and FLT32 which can function as neo-antigens40. For example, NPM1 mutation is reported in up to 60% of cytogenetically normal AML42 where mutation leads to aberrant cytoplasmic expression that is postulated to lead to more efficient HLA presentation43. Our data suggest that rs3916765 affects AML risk by modulating immune recognition of mutated cells, although further work is required to determine which leukemia-specific neo-antigens are involved. The DQB1*03:02-DQA1*03:01 haplotype is associated with increased risk of autoimmune diseases including celiac disease44 and type 1 diabetes46. Reports of concomitant celiac disease and AML are very rare47 consistent with the DQB1*03:02-DQA1*03:01 haplotype having pleiotropic and opposing effects on risk of these two diseases. Taken together, these data implicate dysregulated immune function as a risk modifier for cytogenetically normal AML.We also identify a major susceptibility locus for cytogenetically normal AML at the 6q21.32 cis-regulated AKR1B1 (or related superfamily member AKR1B10) at the 7q33 locus modulates risk of cytogenetically normal AML. AKR1B1 and AKR1B10 encode members of the aldo-ketoreductase superfamily which catalyze the reduction of numerous aldehydes, including the aldehyde form of glucose to generate fructose via the polyol pathway48. Altered glucose metabolism is a hallmark of cancer, where malignant cells switch energy production from mitochondrial oxidative phosphorylation to glycolysis49. In AML, a switch to glycolytic metabolism is associated with disease progression and poor outcome50. Unlike normal monocytes, AML cells can also utilize fructose as an alternative substrate for glycolysis with expression of the GLUT5 fructose transporter as a major regulator of fructose metabolism in leukemic blast cells52, and where high expression is associated with increased proliferation, clonogenicity, migration, and invasion of AML cells51. Interrogation of three independent datasets identified a consistent association between elevated AKR1B1 transcript levels and shorter overall survival in AML53 and serum fructose levels are prognostic in AML51, further implicating this pathway in AML disease progression.Our data suggest that differential expression of 52. Furthermore, the identification of a major AML risk variant at the HLA locus on chromosome 6 implicates altered immune function as etiologically important in AML. Our data supports existing evidence of genetic and biological heterogeneity in AML2 and confirm the need for large collaborative studies to improve statistical power and aid the discovery of sub-type specific genetic risk loci.In summary, our study identifies common susceptibility alleles at four genomic locations that modify AML risk, with evidence of sub-type specific risk loci reflecting the existence of multiple etiological pathways to disease development. Further work is required to decipher the functional basis of these risk loci although our data builds on existing evidence demonstrating a role for aberrant histone modification and altered fructose metabolism in AML pathogenesisN = 529)55, the Eurobank transplant study (N = 70)56, the Leukaemia and Lymphoma Research adult acute Leukaemia population-based case control epidemiology study (N = 222)58, and the institutional hematology biobanks of Newcastle University (N = 48) and University of Chicago (N = 250). Cases were genotyped on the Illumina Omni Express/Omni Express Exome BeadChips. For controls, we used publicly available Illumina Hap550K BeadChip data on 2671 individuals from the British 1958 Birth Cohort59.GWAS1 comprised 1119 AML cases of European ancestry from UK Medical Research Council/National Cancer Research Institute AML clinical trials (N = 120) and King’s College London Medical School (N = 144) with cases recruited from local treatment centers and UK clinical trials. AML cases were also obtained from the Study Alliance Leukemia biobank at Dresden University (N = 208)61, the CALGB 9710 APL clinical trial (N = 101)62, the Newcastle University Haematology Biobank (N = 248), and the Munich Leukemia Laboratory (N = 53) which provides a nationwide diagnostic service in Germany. AML cases were also obtained from treatment centers at the Medical University of Graz (N = 32), University Hospital Rigshospitalet Copenhagen (N = 13), and the Universita Cattolica Sacro Cuore Rome (N = 12). All cases were genotyped on the Illumina Omni Express Exome BeadChip. For controls, we used publicly available Illumina Hap1.2M-Duo data on 2477 individuals from the UK Blood Service Control Group, part of the Wellcome Trust Case Control Consortium 2 controls63.GWAS2 comprised 931 AML cases of European ancestry from the Central England Haemato-Oncology and Oncology Research Bank (N = 82)64, the Centre Hospitalier Universitaire Nice, France (N = 42)65, and the University of London UK (GSE20672) (N = 40). AML cases were also obtained from the Austrian Academy of Sciences (N = 223)66, the Acute Leukaemia French Association Clinical Trials at the University of Lille, France (N = 278)68, the Biobank La Fe at the Hospital Universitari i Politècnic La Fe, Spain (N = 15)69, the German-Austria AML Study Group the German-Austrian AML Study Group , and the Cooperative Health Research in the region of Augsburg (KORA) study (N = 5)73. For controls, we used data from 1612 individuals recruited to KORA study73. All genotyping data for cases and controls were generated on the Affymetrix SNP6.0 Array.GWAS3 comprised 991 AML cases of European ancestry from treatment centers at the Department of Hematology and Oncology of the Medical Faculty Mannheim, University of Heidelberg, Germany (N = 515), treatment centers at the Hematology Division at Semmelweis University, Budapest (N = 202)74 and the UK Biobank (N = 260), a large population-based study conducted in the United Kingdom. For controls, we used 3728 individuals from the UK Biobank. Cases and controls were genotyped on the Affymetrix UK BiLEVE Axiom array or the Affymetrix UK Biobank Axiom array with an equal proportion of cases and controls genotyped on each array.GWAS4 (replication) comprised 977 AML cases of European ancestry from the Central England Haemato-Oncology and Oncology Research Bank  and the AML genome-wide association study in the UK ). AML cases and controls for Samples from the Hungarian AML patients were obtained during the standard diagnostic workup at the Hematology Divisions of the 1st and 3rd Department of Internal Medicine, Semmelweis University, Budapest, following ethical approval from the Local Ethical Committee (REF TUKEB-1552012) and the Hungarian Medical Research Council (REF 45371-2/2016/EKU). Saliva and fibroblast samples from Austrian AML patients were collected at the Division of Hematology, Medical University of Graz, Graz, Austria76. The diagnosis of AML was made in accordance with World Health Organisation guidelines.Collection of patient samples and associated clinico-pathological information was undertaken with written informed consent. All studies were conducted in accordance with the Declaration of Helsinki and received local institutional review board or national research ethics approval (Supplementary Table P < 10−3 in either cases or controls) and with a low call rate (<95%). We also excluded SNPs that showed significant differences (P < 10−3) between genotype batches and with significant differences (P < 0.05) in missingness between cases and controls. Individual samples with a call rate of <95% or with extreme heterozygosity rates (±3 standard deviation from the mean) were also excluded from each GWAS. Individuals were removed such that there were no two individuals with estimated relatedness pihat >0.1875, both within and across GWAS. The individual with the higher call rate was retained unless relatedness was identified between a case and a control, where the case was preferentially retained. Ancestry was assessed using principal component analysis and super-populations from the 1000 genomes project as a reference, with individuals of non-European ancestry excluded based on the first two principal components. In order to minimize any impact of population stratification among the European population we excluded outlying cases and controls identified using principal components 1 and 2 for each GWAS  and the Haplotype Reference Consortium reference haplotype panel (http://www.haplotype-reference-consortium.org/) following pre-phasing using ShapeIT (v2.r790)77. All variants with an imputation info score <0.6 or a minor allele frequency of <0.01 were excluded from subsequent analysis.Genome-wide imputation for each GWAS was performed using the Michigan Imputation Server  in GWAS 2 and GWAS 4. Data on NPM1 and FLT3 somatic mutation status was available for 653 and 865 AML cases, respectively, including 411 and 528 cases of cytogenetically normal AML, respectively. PCR mutation analysis was performed as part of routine diagnostics for NPM1 exon 12 and FLT3 exons 14–15 ; rs3916765 was genotyped in GWAS 4 and imputed in GWAS 1, GWAS 2, and GWAS 3 (info score 0.901–0.995); rs10789158 was imputed in all 4 GWAS studies (info score 0.946–0.9775); and rs17773014 was directly genotyped in GWAS 3 and GWAS 4 and imputed in GWAS 1 and GWAS 2 (info score 0.985–0.993). Fidelity of array genotyping and imputed dosages was confirmed using Sanger sequencing in a subset of AML samples (including samples genotyped on both Illumina and Affymetrix platforms) for each sentinel variant with perfect or very high concordance for all four variants  we identified 1 case with evidence of copy number gain. The susceptibility locus at chromosome 1 does not fall within a region reported to be recurrently somatically deleted or amplified in AML. The association signals at 6p21.32 (rs3916765) and 7q33 (rs17773014) were specific to cytogenetically normal AML and evidence of somatic copy number alterations were visible in 0 and 3 cases, respectively . Specifically, there were three cases with evidence of deletions affecting the chromosome 7 risk locus that were not visible cytogenetically. Furthermore, there was no evidence of copy neutral loss of heterozygosity (>2 Mb) at any of the four AML susceptibility loci reported here. Taken together, these data limit the possibility of differential genotyping in cases and controls due to somatically acquired allelic imbalance.The majority of AML cases were genotyped using DNA extracted from cell/tissue samples (blood and bone marrow) taken during AML remission. A minority of AML cases were genotyped using DNA extracted from tissue samples that include leukemic AML cells. As such, we employed a stringent HWE cut-off  we made use of data from the eQTLGen Consortium (http://www.eqtlgen.org/cis-eqtls.html) for whole blood. Benjamini–Hochberg (BH)-adjusted P values were estimated for each gene annotated to within 1 Mb of the sentinel SNP at each AML association signal. Regions with AML susceptibility variants were annotated for putative functional motifs using data from the ENCODE project81.Imputation of classical 55, Germany61, and Hungary74. Briefly, patients were treated with conventional intensive AML therapy including ara-C, daunorubicin, and best supportive care. A subset of high-risk patients in the German cohort were treated with stem cell transplantation60. Overall survival was defined as the time from diagnosis to the date of last follow-up or death from any cause. Data on relapse-free survival was available on 358 AML patients, which was defined as the time from date of first remission to the date of last follow-up in remission or date of AML relapse. Cox regression analysis was used to estimate allele-specific hazard ratios and 95% confidence intervals for each study in analyses that included all AML cases (N = 767) and cytogenetically normal AML (N = 358).The relationship between AML risk variants and survival was evaluated in a total of 767 AML patients (excluding acute promyelocytic leukemia) from the UKFurther information on research design is available in the Supplementary InformationPeer Review FileReporting summary


Consequently, the MAM is suggested to be involved in, and as a possible therapeutic target for, some common diseases and impairment in skeletal muscle function, such as insulin resistance and diabetes, obesity, neurodegenerative diseases, Duchenne muscular dystrophy, age-related muscle atrophy, and exercise-induced muscle damage. In the past decade, evidence suggests that alterations in Ca2+ transport from the ER to mitochondria, mediated by the macromolecular complex formed by IP3R, Grp75, and VDAC1, may be a universal mechanism for how ER-mitochondria cross-talk is involved in different physiological/pathological conditions mentioned above. A better understanding of the ER (or sarcoplasmic reticulum in muscle)-mitochondria Ca2+ transport system may provide a new perspective for exploring the mechanism of how the MAM is involved in the pathology of diseases and skeletal muscle dysfunction. This review provides a summary of recent research findings in this area.The physical contact site between a mitochondrion and endoplasmic reticulum (ER), named the mitochondria-associated membrane (MAM), has emerged as a fundamental platform for regulating the functions of the two organelles and several cellular processes. This includes Ca Sarcoplasmic reticulum (SR) is a special form of ER in striated muscle; it is specialised in the lipid synthesis, Ca2+ modulation, and excitation–contraction coupling. Structurally and functionally, these two organelles are connected via the mitochondria-associated membrane (MAM), a contact site that acts as a platform for inter-organelle communication  variations on either the outer mitochondrial membrane (OMM) surface or within mitochondria ([Ca2+]mt), in response to stimulation of the IP3R, are frequently measured for exploring the Ca2+ communication between the ER and mitochondria in the liver and brain mt, i.e., mitochondrial Ca2+ overload, could cause a constant opening of the mitochondrial permeability transition pore (mPTP). This may subsequently lead to mitochondrial swelling, rupture of the OMM, and the collapse of the mitochondrial membrane potential, resulting in structural and functional damage to mitochondria  in the mitochondrial intermembrane space is indistinguishable from that in the cytoplasm ([Ca2+]c) (around 100 nM)  under resting conditions or even in response to physiological stimulation (approximately 1 μM)  at the ER–mitochondrial interface is much higher than that in bulk cytoplasm. This unique [Ca2+] localisation has been directly demonstrated by monitoring the local [Ca2+] at the ER–mitochondrial interface by fluorescent interorganelle linkers  in local regions near the mitochondrial surface is 5–10-fold higher than that in bulk cytoplasm [2+] at the ER–mitochondrial interface may then rise to several tens of micromolar during the activation of the IP3R. There are two important types of Ca2+ release channels at the ER membrane, ryanodine receptor (RyR) and IP3R. While IP3R is mainly for forming the high Ca2+ microdomain mt, and at times activation of this protein can result in [Ca2+]mt 20 times higher than the global increase in [Ca2+]c [3R may evoke a large [Ca2+] change at the ER–mitochondrial interface even when the increase in [Ca2+]c is suppressed in bulk by the ethylene glycol-bis(β-aminoethyl ether)-N,N,N’,N’-tetraacetic acid (EGTA)/Ca2+ buffer c . Additio+ buffer . These fchondria . Moreoveof Grp75 . The Grpof Grp75 . This prnd VDAC1 .2+ transport, with the [Ca2+]mt being increased by moving the ER closer to the mitochondria mt with the [Ca2+]c in response to caffeine when using a fast Ca2+ buffer BAPTA ethane-N, N, Nʹ, Nʹ-tetraacetic acid) in the cytosol, with the mitochondrial Ca2+ concentration increasing and cytosolic Ca2+ showing little change mt was found to be increased, but [Ca2+]c showed no change, with an accompanying rise in MCU expression and activity c and [Ca2+]mt [3R expression does not enhance mitochondrial Ca2+ uptake because of the reduction of the complex quantity of IP3R-Grp75-VDAC1 [2+ transport is a suspected contributor to the reduced [Ca2+]mt in DMD [Muscle weakness and wasting have long been confirmed as pathogenic DMD symptoms . It is aystrophy . An incr[Ca2+]mt . However75-VDAC1 . Thus, tt in DMD . 2+ transport may also potentially play a role in muscle dysfunction in ageing through mitochondrial Ca2+ homeostasis impairment. Further, tetanic stimulation on the flexor digitorum brevis of mice was reportedly the cause of reduced mitochondrial Ca2+ uptake, along with a decrease in the association between mitochondria and CRUs in aged mice mt caused by the increased MCU expression [2+ homeostasis in the SR and mitochondria are said to cause the functional decline of skeletal muscle in ageing. The defective SR-mitochondria Ca2+ cross-talk may precede muscle function impairment in age-related dystrophy, but further studies on molecular mechanism and Ca2+ signalling are required to understand this explanation better.Disrupted SR-mitochondria Caged mice . In agedged mice . Gill etged mice . An amela2+ load . Exercispression , which ipression . Based o2+ release through Ca2+ channels is an essential cause of ER stress, and ER stress can be attenuated by the cone-specific gene deletion of IP3R1 [2+ homeostasis, mostly through mitochondrial Ca2+ overload, also occurs in the diseases mentioned above, and it is probably due to the increased ER-mitochondria Ca2+ transport. In the early phase of ER stress, mitochondria were found to migrate to the ER’s periphery [2+ overload was shown to be caused by mitochondrial migration to the ER surface, resulting in mitochondria-dependent apoptosis [3R-Grp75-VDAC1, Ca2+ transport from the ER to the mitochondria is likely to contribute to podocyte apoptosis by facilitating mitochondrial Ca2+ overload [2+ transport from the ER to the mitochondria may contribute to the ER stress and mitochondrial Ca2+ overload at the same time, but this speculation needs further experimental evidence. It is commonly accepted that ER stress, which can be caused by ageing, genetic mutations, or environmental factors, can result in various diseases, including those mentioned above . The incof IP3R1 . The diseriphery ,87, whilpoptosis . In addioverload . As can 3R-Grp75-VDAC1 pathway, in calcium ion transport from the endoplasmic/sarcoplasmic reticulum to mitochondria, in relation to normal and abnormal cellular functions. Current evidence suggests that the distance between the ER/SR and mitochondria, and the modulation of the IP3R, Grp75, and VDAC1 activities, play a critical role in regulating mitochondrial Ca2+ homeostasis. A disruption in this pathway and subsequently Ca2+ homeostasis  is believed to be a common causative factor in some health conditions, such as neurodegenerative diseases, obesity, insulin resistance, cancer, and ischemia/reperfusion damage, as well as in age-related muscle dystrophy and intensive exercise-induced reduction of skeletal muscle function. However, the exact cause-and-effect relationship and the potential physiological/pathological mechanisms under specific conditions require further investigation to shed light on potential therapeutic targets or interventions for such conditions. This review focused on the mitochondria-associated membrane’s role, particularly the IP


As a new type of non-coding RNA, the role of circular RNA (circRNA) in various diseases and tumors has received considerable attention. Studies have shown that circRNAs play an important role in the progression of acute myeloid leukemia (AML) via different mechanisms. However, the specific underlying molecular mechanism of circRNAs in the proliferation of AML cells remians unclear. This study aimed to clarify the biological role and mechanism of circCRKL in AML. The results indicated low circCRKL expression in AML cell lines and samples. Moreover, the overexpression of circCRKL inhibited the proliferation and colony-forming ability of AML cells, while its silencing promoted them. In addition, bioinformatics tools and luciferase assays revealed that circCRKL could sponge miR-196a-5p and miR-196b-5p to promote the expression of p27. Furthermore, circCRKL inhibited AML cell proliferation via the miR-196a-5p/miR-196b-5p/p27 axis, suggesting a potential new target for AML therapy. Acute myeloid leukemia (AML) is a heterogeneous and invasive hematological malignancy . Over thCircular RNAs (circRNAs), a new type of non-coding RNAs, exhibit continuous closed loops and stable expression in mammalian cells while reportedly participating in the progression of many diseases by regulating various biological processes, including gene expression and transcription . GeneralV-Crk Avian Sarcoma Virus CT10 Oncogene Homolog-Like (CRKL) GeneID:1399) gene is reported to be involved in the progression of myeloid leukemia ,13, whet99 gene iThis study aimed to identify a new circRNA/miRNA/mRNA network in AML. Here, we hypothesized that circCRKL suppressed the proliferation of AML cells via miR-196a-5p/miR-196b-5p/p27 axis. CircCRKL may be a promising target in the treatment of AML.Bone marrow (BM) samples were obtained from 34 primary AML (non-acute promyelocytic leukemia) patients and 34 healthy volunteers from the Institute of Hematology of Union Hospital affiliated of Tongji Medical College. The detailed clinical parameters of the AML samples are shown in Table S1. All samples were obtained with the informed consent of the patients and were approved by Union Hospital, Tongji Medical College, Huazhong University of Science and Technology Ethics Committee ([2020]0075–01).2.Human AML cell lines, KG-1a (French American British classification systems-M0 (FAB-M0)), KG-1 (FAB-M1), Kasumi-1 (FAB-M2), and the human BM stromal cell line, HS‐5, were purchased from the American Type Culture Collection . THP1 (FAB-M5) and MOLM13 (FAB-M5)  were obt−ΔΔCt). The primer sequences are provided in Table S2.The total RNA was isolated using Trizol reagent . RNase R treatment was processed at 37°C with RNase R . Complementary DNA (cDNA) was then directly synthesized and reversed with PrimeScript RT Master Mix . qRT-PCR was performed using the SYBR kit  and analyzed via a StepOne Plus system  . GAPDH aThe pCD5‐ciR vector  was used for circCRKL overexpression. The human p27 cDNA was synthesized and inserted into pcDNA3.1 by Vigene . The shRNAs against circCRKL and p27 were designed and cloned into pGPU6-GFP-Neo . The miR-196a-5p/miR-196b-5p mimics, negative control (miR-NC), and anti-miR-196a-5p/anti-miR-196b-5p inhibitor were purchased from RiboBio . The transfection was performed with lipofectamine 3000 reagent  as previously described . The det3 cells per well were plated into a 96-well plate  and incubated at 37°C. After incubation with 10 μL of CCK-8 reagent  for 2 h, the cell viability was measured at 450 nm using a microtiter plate reader [Approximately 5 × 10e reader .6) were fixed using 70% cold absolute ethyl alcohol, stained with propidium iodide reagent , and analyzed using flow cytometry (BD LSRFortessaTM X-20) [AML cells (1 × 10TM X-20) . ModFit 4) were suspended in 1 ml of 0.7% agarose gel mixed with cell culture medium and added to the bottom layer. The surrounding wells were supplemented with 1 ml of phosphate buffered saline  and placed in an incubator after the solidification of the gel. The clones were stained with MTT -2,5-diphenyltetrazolium bromide)  and scored after 10–14 days [The 6-well culture plates were pre-coated with 2 ml of 1.2% agarose gel  mixed with cell culture medium (bottom layer). Then the AML cells (0.3 × 10–14 days .The transfected AML cells were lysed with Radio-Immunoprecipitation Assay (RIPA) buffer  containing magnetic beads conjugated with anti-Ago2 antibody . A circRIP assay was conducted as previously described . Biotin-The wild-type (wt) and mutated (mut) sequences of the circCRKL and CDKN1B 3ʹ untranslated regions (UTR) containing miRNA binding regions were amplified and cloned into a pmirGlo vector . A dual-luciferase assay was conducted as described .The total proteins were extracted with RIPA lysis buffer, while bicinchoninic acid  was used to detect the protein concentration. Western blotting analysis was performed as previously described , using pP < 0.05 was accepted as statistically significant.An unpaired Student’s t-test was used to evaluate the group differences between normally distributed variables, while one-way analysis of variance was used for comparisons between three or more groups. The data were expressed as the mean ± standard deviation using GraphPad Prism 8.01  and SPSS 19.0 statistical software. This study hypothesized that circCRKL inhibited AML cells proliferation and demonstrated that circCRKL was down-regulated in the AML samples and cell lines using qRT-PCR. A CCK8 assay, a colony-forming assay and cell cycle analysis were used to reveal the role of circCRKL in proliferation of AML cells. The relationship between circCRKL, p27, and miR-196a-5p/miR-196b-5p was predicted using bioinformatics tools and verified via luciferase assays. Furthermore, this study revealed that circCRKL inhibited AML cells proliferation by sponging miR-196a-5p/miR-196b-5p to affect p27 expression.The expression of circCRKL in 34 clinical AML samples and normal samples (n = 34) was determined using qRT-PCR to assess the role of circCRKL in AML.  shows thCircCRKL was overexpressed in THP1 cells and downregulated in MOLM13 cells to explore its role in AML cells . The CCKStudies have shown that sponge miRNAs facilitate circRNA functionality and that circCRKL is mainly distributed in the cytoplasm. Therefore, an RIP assay was performed to elucidate whether circCRKL acted as a miRNA sponge to enable its functionality. The results showed that the circCRKL was significantly enriched by the AGO2 antibody , indicatTwo bioinformatics algorithms, namely TargetScan  and miRDAML is considered a deadly hematopoietic malignancy. Despite advances in AML therapeutic regimens, approximately half of patients experience recurrence or resistance and die of the disease. Therefore, the identification of new therapeutic targets is urgently needed. With the emergence and development of high-throughput sequencing technologies, more and more circRNAs have been found to be abnormally expressed in carcinomas and other diseases . In addiSome circRNAs contain miRNA response elements that can be used as ceRNA to sponge miRNAs, regulating downstream gene expression and affecting the biological functionality of diseases. For example, circ_0001162 is involved glioma development via the miR-936/ERBB4 axis . CircRNAP27, encoded by the CDKN1B gene, is a member of the kinase inhibitory protein (Kip) family and a cyclin-dependent kinase inhibitor that mediates cell-cycle inhibition. The CDKN1B gene encoding human p27 is involved in regulating the cyclin E/cyclin-dependent kinase 2 (CDK2) complex . Howeverin vivo experiments are necessary to support the conclusions of this study.However, this research still presents some limitations. According to reports, p27 is also involved in tumorigenesis via the independent cyclin-cdk pathway . StudiesThis work demonstrates that circCRKL is down-regulated in AML, showing potential as a therapeutic target for AML, while it can sponge miR-196a-5p and miR-196b-5p to inhibit p27 expression. This research explored the potential mechanism of circCRKL functionality in AML cells and provided a potential target for AML treatment.Click here for additional data file.


Myocardial ischemia-reperfusion injury (MIRI) is common in patients with acute coronary syndrome (ACS) after PCI treatment, which seriously affects the efficacy of revascularization and hinders the postoperative recovery of patients; therefore, the current study is focused on determining effective methods in the treatment of MIRI. Antiplatelet therapy is a routine treatment for ACS, and its benefits for treating MIRI have been previously verified. With the development of traditional Chinese medicine (TCM), many TCM preparations are widely used in the clinic. Many basic and clinical studies have shown that TCM can be used together with antiplatelet drugs, and the safety and efficacy when TCM is included in the treatment are better than when antiplatelet drugs are used alone. This paper summarizes the current research progress of traditional Chinese medicine and Western medicine in the treatment of MIRI to provide a theoretical basis for further research and clinical treatment. Acute coronary syndrome (ACS) is one of the leading causes of death worldwide . ACS occPlatelets are discoid anucleate hematopoietic cells with an average diameter of 1.5–3.0 mm. They are produced by megakaryocytes in the bone marrow, circulate in blood, and are cleared by the reticuloendothelial system . These cSalvia miltiorrhiza, Ligusticum chuanxiong, and safflower. Currently, TCM and Western medicine have increased the understanding of antiplatelet therapy. This paper summarizes the current clinical research in both TCM and Western medicine for the treatment of MIRI and provides a theoretical basis for further clinical research in the future.From the philosophy of TCM, there is a deficiency of qi in the pathogenesis of MIRI, and this can lead to the development of blood stasis, and blood stasis further aggravates qi deficiency; therefore, blood stasis is one of the important pathological processes in MIRI . Many baBlocking platelet aggregation during PCI in patients with an acute myocardial infarction (AMI) is the standard treatment for inhibiting intravascular coagulation and reducing stent rethrombosis. Aggregates of platelets, leukocytes, and other substances produced after a coronary endothelial injury can occlude microcirculatory vessels, causing MIRI in ACS patients undergoing PCI. Platelets are also the first blood cells to respond to MIRI. Therefore, the effect of antiplatelet therapy on MIRI after PCI has been a research focus. Platelets have multiple pathophysiological effects in response to MIRI.One of the effects is platelet microthrombosis . Some reIn addition, platelet aggregation is an early marker of AMI and is also related to cardiac no-reflow in patients with STEMI after PCI , 19. RenAggregation of platelets with leukocytes is also an important mechanism of MIRI, and it has been demonstrated in animal models and clinical trials that MIRI can induce an increase in platelet neutrophil complexes –21. StudPlatelets are activated and play an important role after MIRI stimulation, platelet activation causes an increase in the mean platelet volume and an increase in the secretion of microvesicles (MVSs), apoptotic bodies, and exosomes, and various mediators carried by these extracellular vesicles, such as proteins, mRNAs, and microRNAs, are released , 25. ActTherefore, with the improved understanding of MIRI, it is recognized that platelets play important roles in the pathophysiology of MIRI and that antiplatelet therapy can inhibit the aggregation of platelets within the myocardium and inhibit the release of vasoconstrictive substances after platelet activation, indicating its great potential for MIRI therapy and that it is an important method for the treatment of MIRI in modern medicine.Antiplatelet drugs are usually divided into anti-thromboxane A2, ADP receptor antagonists, PDE3 inhibitors, platelet GP IIb/IIIa receptor antagonists, and so on.Dual antiplatelet therapy (DAPT) is the cornerstone of the treatment of ACS. Aspirin and clopidogrel are the most widely used antiplatelet drugs in the world. The CURE trial  evaluateGuidelines , 4 recomThe choice of P2Y12 receptor antagonists was based on clinical trial selection. The PLATO trial  treated MIRI is an important factor affecting the expansion of myocardial infarct size. DAPT may reduce myocardial infarct size by treating MIRI. Other antiplatelet agents can also be combined with DAPT to prevent against myocardial injury, such as GP IIb/IIIa inhibitors. Three GP IIb/IIa inhibitors are abciximab, tirofiban, and eptifibatide. Among them, abciximab is the most studied GP IIb/IIa inhibitor . Basic rA meta-analysis evaluated the safety and efficacy of GP IIb/IIIa inhibitors during PCI. Forty-eight studies involved more than 33,000 patients. In short, the use of GP IIb/IIa inhibitors after PCI has been shown to be beneficial, as shown by a reduction in the 30-day mortality, nonfatal myocardial infarction, and emergency revascularization. However, this benefit is accompanied by a substantial risk of bleeding. The benefit disappeared 6 months after treatment . OverallCurrently, 12 months of DAPT is commonly used in the clinic to prevent MIRI after PCI in patients with ACS. There are meta-analyses  which inSalviae miltiorrhizae, Panax notoginseng, and borneol are the main components of CDDP, in which Salvia miltiorrhiza can activate blood circulation, remove blood stasis, and improve myocardial blood supply. Panax notoginseng can stop bleeding, invigorate blood circulation, remove blood stasis, and relieve pain. Borneol reduces swelling and relieves pain.Compound Danshen Dripping Pill (CDDP) is a proprietary Chinese medicine based on TCM theory and modern medical technology. It regulates qi, relieves pain, promotes blood circulation, and removes blood stasis. Radix Panax notoginseng saponins [P < 0.00001). Compared with PCI alone, CDDP combined with PCI significantly reduced WBV, PV, and EAI (P < 0.00001), suggesting that CDDP can reduce the risk of postoperative PCI and treat MIRI by inhibiting platelet thrombosis and anticoagulation.From the point of view of modern pharmacology, CDDP has the effects of antiplatelet aggregation and antithrombosis, reducing blood viscosity, protecting against reperfusion injury, and improving microcirculation. The main active components are salvianolic acid B and saponins . A recensaponins  includedP < 0.05). It is suggested that CDDP can significantly improve the myocardial microcirculation in patients with no reflow after PCI. DAPT combined with antithrombotic therapy is often adopted in the perioperative period after PCI of acute MI to prevent MIRI. The clinical trial designed by Wei et al. [A meta-analysis  includini et al.  includedClinical studies  have fouNaoxintong capsules are based on the theory of TCM, which are made from the Buyang Huanwu decoction. It has the effect of replenishing qi and activating blood circulation, removing blood stasis, and dredging collaterals. Buyang Huanwu decoction is a clinically recognized prescription for the treatment of CHD with qi deficiency, blood stasis syndrome, and cerebrovascular disease .β and NLRP3 and improve cardiac function in mice with MIRI. There are also basic studies [Naoxintong capsules can reduce myocardial injury in patients with CHD. It has been found that taking Naoxintong capsules one week before PCI can reduce the levels of serum hs-CRP, CK-MB, and cTnI after PCI, reduce the myocardial injury caused by PCI, and reduce the damage caused by MIRI . There m studies  that NaoHuan  discusseClinical studies have found that the combination of Naoxintong capsules with aspirin can provide optimized antiplatelet effects in patients with cardiovascular and cerebrovascular diseases. The study  includedThere are many mechanisms of Naoxintong in the treatment of MIRI, such as anti-inflammatory effects, antiplatelet aggregation, and improvement of vascular endothelial function that may be involved in the treatment of MIRI , but preSalvia miltiorrhiza and safflower, which has the effect of promoting blood circulation, removing blood stasis, and dredging meridians and collaterals. It mainly contains acids, tanshinones, flavonoids, and other chemical components [Danhong injection (DHI) is composed of mponents . By compmponents , it was In the theory of TCM, the pathogenesis of MIRI is a blood stasis block. DHI has the effect of promoting blood circulation and removing blood stasis, while the aforementioned methods have been shown to be effective treatments for MIRI. Basic studies have found that DHI can increase the maximum platelet aggregation time of rabbits, effectively reduce the rate of platelet aggregation, and inhibit thrombosis in rabbits . DHI canP < 0.05), and inflammatory factors and vascular endothelial function were improved to some extent. DHI can also interact with aspirin, and the combination of drugs can inhibit the activity of aspirin esterase, enhance the antiplatelet effect of aspirin, increase the serum concentration of salicylic acid, and enhance the anti-inflammatory activity of salicylic acid [DHI is often used in combination with antiplatelet drugs in the clinic. In a clinical study , DHI comlic acid . NetworkSalvia miltiorrhiza. It dilates the coronary arteries, promotes the scavenging of oxygen free radicals, improves microcirculation, and protects the myocardium. Clinically, it is often used in combination with antiplatelet drugs such as aspirin and clopidogrel in the treatment of cardiovascular diseases.Sodium tanshinone IIA sulfonate injection (STS) is prepared by the sulfonation of fat-soluble tanshinone IIA, which is the main active component of Studies in vitro and in vivo have confirmed that the pharmacological effects of STS include antilipid peroxidation to prevent MIRI from preventing platelet aggregation and improving hemorheology . Maione P < 0.05), and the postoperative MIRI was improved to some extent [STS is widely used in clinics. In clinical studies , STS come extent .The study  includedMIRI is an important sequela affecting the curative effect after revascularization in patients with ACS. Platelet aggregation or platelet activation is involved in the development of MIRI. Antiplatelet therapy with Western medicine, as the standard treatment after revascularization, can inhibit platelet aggregation and platelet activation. Many studies have confirmed its benefit in MIR therapy. At present, aspirin combined with P2Y12 receptor antagonists is the main antiplatelet therapy. With the improvement in research, there are more P2Y12 receptor antagonists to choose from, and there are new antiplatelet drugs to supplement DAPT, but the clinical benefits for MIRI are usually accompanied by an increase in the risk of bleeding. Some studies also believe that there is no significant improvement in clinical symptoms. TCM preparations, guided by theories such as invigorating blood circulation and eliminating stasis in TCM, are usually combined with Western antiplatelet therapy in the clinic, which is more effective clinically than antiplatelet agents alone, probably through the interaction with antiplatelet agents, and some studies considered that it does not increase the risk of bleeding. The combination of TCM and Western medicine in the treatment of MIRI is promising with regard to optimizing the antiplatelet treatment regimen. Millions of patients currently accept the treatment of integrative medicine in China, and the effectiveness and safety of these treatments have been fully verified in the clinic. Further clinical and basic research is needed to clarify these mechanisms and the effectiveness and safety of the drugs.


This cross-sectional study examines variation in SARS-CoV-2 infection risk and socioeconomic disadvantage among a Mayan-Latinx population in the Fruitvale, California, community. Free SARS-CoV-2 testing was provided for individuals of all ages on September 26 to 27, 2020. Adults gave verbal consent for themselves and for participating children. Anterior nasal swab samples were obtained for polymerase chain reaction (PCR) testing to detect the virus, and venous blood was collected to detect immunoglobin G antinucleocapsid antibodies. Adults completed a survey on sociodemographic characteristics at the testing event. Interviewers fluent in Spanish, 2 Mayan languages, and more than 4 other languages were available. We analyzed cross-sectional associations between demographic and socioeconomic indicators and SARS CoV-2 infection using χ2 tests and logistic regression analyses adjusted for age and sex and accounting for household clustering. Statistical significance was set at P < .05. Data analyses were conducted in Stata version 16 (StataCorp). Additional information regarding the methods appear in the eAppendix in the This cross-sectional study was approved by the UCSF institutional review board and followed the Strengthening the Reporting of Observational Studies in Epidemiology (P < .005), report food insecurity , have difficulty finding work due to the pandemic , lack a regular medical practitioner , and have no health insurance  (P = .04); 41 (44.6%) spoke a Mayan language at home. Mayan and non-Mayan Latinx participants had significantly greater odds of having a positive PCR test compared with non-Latinx participants . Mayan individuals were significantly more likely to have positive serology results compared with non-Latinx participants  ; 108 (9.1%) were Mayan individuals, 661 (55.7%) non-Mayan Latinx individuals, and 417 (35.2%) non-Latinx individuals. Compared with other Latinx individuals, Mayan individuals were more likely to live in households with 5 or more people  . Mayan i < .001) .1 Our data highlight heterogeneity within the Latinx community, with Mayan individuals having even higher risk than other Latinx individuals. Findings related to socioeconomic disadvantage, including large household size, low income, and food insecurity, likely reflect the heightened susceptibility of Mayan individuals to the pandemic.6 In addition, limited English proficiency and access to health care pose challenges for effective public health messaging.We found that Latinx participants were more likely to have current SARS-CoV-2 infection than non-Latinx participants, reflecting state and national trends.Limitations that reduce generalizability include that the study analyzed a convenience sample of those seeking testing, and that testing and medical referrals were prioritized over questionnaire completion, resulting in missing data. Unmeasured confounding factors could attenuate results.This study underscores the need to consider heterogeneity within Latinx communities and to prioritize subgroups with higher risks, such as Mayan individuals, in health policies and outreach. Limited Spanish and English proficiency reduce this population’s access to health information and care. Few CBOs or public health departments have Mayan language speakers to provide information and perform contact tracing. Those that do are underresourced. Understanding differential risks within the heterogenous Latinx population can guide more efficient targeting of services. Failure to engage with communities with higher risks increases the likelihood of ongoing transmission and may hinder SARS CoV-2 vaccine uptake.