As one of the common cardiovascular diseases, acute myocardial infarction (AMI) is characterized by a high mortality rate, frequent complications, and a serious threat to human health and quality of life. Traditional Chinese medicine injection (TCMI) has been used clinically to treat AMI; however, there is no uniform standard for clinical treatment of AMI. The purpose of this study is to evaluate the efficacy and safety of different TCMI by using systematic review and network meta-analysis.According to the strategy, the authors will retrieve both 4 Chinese databases and 3 English databases by June 30, 2020. After a series of screening, randomized controlled trials will be included related to TCMI for AMI. Two researchers will use Aggregate Data Drug Information System and STATA 15.0 to analyze the data. Finally, the evidence grade of the results will be evaluated.This study will provide a reliable evidence for the selection of TCMI therapies for AMI.The results of this study will provide references for evaluating the influence of different TCMI therapies for AMI, and provide decision-making references for clinical research.DOI 10.17605/OSF.IO/FYGBT. There are 5 types: Type 1 AMI caused by acute atherosclerotic thromboembolic coronary events; Type 2 AMI is caused by increased oxygen demand or decreased oxygen supply and may occur in patients with or without significant coronary artery disease; Type 3 AMI is only applicable to rare cases of death in patients with symptoms suggestive of myocardial ischemia. It can also be presumed to be newly occurred, ST-segment elevation, or new left bundle branch block. Type 4 and type 5 AMI are iatrogenic acute myocardial infarction associated with percutaneous coronary intervention (PCI) and coronary artery bypass grafting, respectively. However, in clinical practice, it is difficult to distinguish type 2 AMI from type 1 AMI, and it is also difficult to distinguish type 2 AMI from other myocardial injury or nonischemic diseases associated with elevated troponin, such as myocarditis, Takotsubo cardiomyopathy, or septic shock. Therefore, the prevalence of type 2 AMI varies greatly, ranging from 1.6% to 29.6%.–7 The pathological factors of myocardial ischemia and infarction are various, including regional myocardial ischemia, global ischemia, myocardial shock, hibernating myocardium (chronic coronary insufficiency), myocardial necrosis, or fibrosis.–11 Myocardial ischemia will seriously impair the systolic and diastolic functions of the heart, and patients with AMI are prone to irritability and fear, thus making sympathetic nerves excited, catechol amine level increased, myocardial load increased, and the disease worsened. Complications of AMI include various forms of hemodynamic insufficiency, such as acute heart failure, left ventricular remodeling, chronic heart failure, and cardiogenic shock.,15Acute myocardial infarction (AMI) is caused by acute blockage of coronary arteries and interruption of blood flow, which causes continuous ischemia and hypoxia in myocardial cells. PCI operation is the preferred method in the treatment of AMI. However, the phenomenon of "no reflow (NR)" may be caused after surgery, the incidence ranged from 5% ∼ 50%,,19 may lead to myocardial infarction area increases, affect heart function recovery, thereby increasing the forward end point events, lead to a significant portion of patients cannot obtain a satisfactory response to treatment. Therefore, the treatment of AMI is in urgent need of better treatment strategies.At present, the typical treatment of AMI patients is based on the main principles of opening infarcted blood vessels as soon as possible, promoting myocardial ischemia area reperfusion, saving dying myocardium, and actively managing complications.,22 At present, the main traditional Chinese medicine injection (TCMI) treatments for AMI include Shenmai injection, Shenfu injection, Xinmailong injection, Shengmai injection, Astragalus injection, etc. For example, shenmai injection can significantly reduce the ischemic myocardial area and increase the ejection fraction during early thrombolysis of AMI. Danhong injection (DHI) can reduce ST segment elevation myocardial infarction (STEMI) patients with high risk of reflow.Existing evidence shows that traditional Chinese medicine (TCM) has gained rich experience in the treatment of acute myocardial infarction, and the combination of TCM and Western medicine (WM) can further improve the prognosis of patients.At present, some studies have systematically evaluated the efficacy of TCMI in treating AMI. However, there is no network meta-analysis (NMA) on the differences between different types of TCMI in the treatment of AMI. The purpose of this study was to evaluate the efficacy and safety of different TCMI and to provide reference and evidence for clinical application.22.1https://osf.io/fygbt), registration number: DOI 10.17605/ OSf.io /FYGBT. This protocol follows the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Protocols guidelines.The NMA protocol has been registered on the Open Science Framework platform , intervention (I), comparator (C), outcome (O), and study design (S).2.3.1All studies including patients with AMI diagnosed by any set of criteria were eligible for inclusion, such as 2012 ESC Guidelines on acute myocardial infarction (STEMI), Heart Disease and Stroke Statistics—2019 Update: A Report From the American Heart Association. Patients with other serious complications were not included, and the course and severity of the disease were approximately the same regardless of sex, age, nationality, or educational background.2.3.2Treatment group TCMI in combination with WM or taking TCMI alone, the control group didn’t intervene, placebo or WM. TCMI include Shenmai injection (SMI), Xinmai long injection (XMLI), Yiqi Fumai injection (YQFMI), Shenfu injection (SFI), DHI, Huangqi injection (HQI), Shenqi Fuzheng injection (SQFZI), etc.2.3.3Primary outcomesLeft ventricular ejection fraction (LVEF);N-terminal forebrain natriuretic peptide (NT-proBNP);Major cardiovascular adverse events (MACE).Secondary outcomesST segment regression degree;Venous thrombolysis rate;Serum inflammatory factors include high sensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α);Adverse reaction (AR).2.3.4This study is a systematic review with NMA of randomized controlled trials (RCTs) on the TCMIs for the AMI. All relevant RCTs using TCMIs for the AMI will be included. Quasi-RCTs, duplications, review documents, animal trails, clinical experience, and case reports will be excluded. Additionally, only Chinese and English literature will be searched for this study.2.4Three English databases including Medline, The Cochrane Library, Embase and 4 Chinese databases were searched, including China Knowledge Infrastructure Database, China Biomedical Science, Chongqing VIP, and Wan-Fang. The time limit for literature retrieval is from the establishment of each database to 30 June 2020. The language is limited to English and Chinese. To obtain the potential nonelectronic literature, relevant magazines and medical journals will also be filtered for further search. In these databases, strict restrictions will be placed to exclude the types of studies that are not RCTs. Chinese search terms mainly include: “acute myocardial infarction;” English search words include “acute myocardial infarction,” “AMI,” “injection,” “Traditional Chinese medicine,” “Chinese herbal medicine.” Taking Medline as an example, the initial retrieval strategy is shown in Table 2.5https://endnote.com), the duplicate literature was eliminated; Conduct a preliminary screening by reading the headline summary to exclude literature that does not meet the inclusion criteria; Reading the full text and making final selections; Data extraction using a predesigned data extraction table for the included literature and cross-checking the results; In case of disagreement, the third researcher Qingjuan Wu will be called upon to assist in judgment. Microsoft Excel  will be used to extract the data and collecting relevant information. Data extraction mainly included basic information of the literature , basic information of study subjects  in Cochrane Handbook 5.1.0. Two trained researchers Wei Zhao and Qingjuan Wu will independently evaluate the risk of bias of the included studies. In case of dispute, submit to corresponding author Jun Li for arbitration. random Sequence Generation, allocation Concealment possibility; blinding of participants and personnel; blinding of outcome assessment; incomplete outcome data; selective reporting; other bias. Based on the above 7 entries, the judgment is divided into 3 levels: low risk of bias, high risk of bias, and unclear risk of bias.Cochrane bias risk assessment tool will be used to assess the risk of RCTs being included in NMA, 7 items are included:2.72.7.1 and uses Markov chain-Monte Carlo algorithm to make Bayesian inference. Iteration operations were performed according to the following preset model parameters: 4 chains were used for simulation analysis, with initial value of 2.5, a step size of 10, annealing times of 20,000 and 50,000 simulation iteration times. ADDIS software is used to draw network evidence diagrams of different outcome indicators, and odds ratio or standardized mean differences is used for statistical analysis, both with 95% credible intervals. According to the results of the NMA, rank probability plot of various TCMI therapies is generated and sorted by dominance, with Rank1 being the optimal sort.This study uses Aggregate Data Drug Information System (ADDIS) 1.16.8 for NMA,2.7.2P > .5), the consistency model was used for analysis. On the contrary, inconsistent model is adopted for analysis. If the consistency model is adopted, then the stability of the results is verified by the inconsistency model: When the inconsistency factors including 0, at the same time inconsistency standard deviation including 1 says the result of inconsistency model is more stable and reliable. At the same time, the preset parameters are used for iterative operation, and the convergence degree of iteration is judged by the potential scale reduced factor (PSRF). When the PSRF value is close to or equal to 1 (1≤PSRF≤1.05), the convergence is complete, indicating good stability of the model and reliable analysis results. If the PSRF is not in this range (1≤PSRF≤1.05), the iteration will continue manually until the PSRF value reaches the range standard.In this study, node-split model was used to analyze the consistency of data. When the statistical difference was compared directly and indirectly (2.7.3https://www.stata.com). Evaluate the heterogeneity between studies through I2. When I2 > 50%, it indicates that the heterogeneity between studies is large, using a random effect model; when I2 < 50%, it indicates that the heterogeneity between studies is small or there is no difference qualitative, using a fixed effect model. When the heterogeneity is greater, the source of heterogeneity should be further sought.Before the combination of effect size, the heterogeneity of the included literature is tested using STATA 15.0 software (Stata Corp, College Station, TX, 2.7.4If necessary, sensitivity analysis will be used to assess the impact of the studies on the random effects model. After each study was excluded one by one, the data analysis was carried out again to determine the stability of the results. If there is no qualitative change in the combined effect shown in the results, the results are stable.2.7.5If there is clinical and methodological heterogeneity, we will conduct a subgroup analysis of the patient's age, onset of AMI, duration of treatment, or study quality.2.7.6If 10 or more studies are included in the NMA, a comparison-adjusted funnel plot is developed using Stata to evaluate the presence of small sample effects or publication bias in the intervention network. If the plot is asymmetric and there is no inverted funnel shape, it indicates that there may be publication bias. The reasons may be related to the small sample size, allocation concealment, and insufficient implementation of blind method.2.7.7If the literature information is clearly incorrect or incomplete, we will contact the first author or the first author of the literature via email address. If no response is received, the document should be deleted.2.7.8 Based on bias, inconsistent, inaccurate, indirect, and the risk of publication bias 5 degradation factors, the quality classification for the 4 level of evidence: high, medium, low, and very low.To grade evidence quality and understand the current situation of evidence rating thereby analyzing possible problems, the Grading of Recommendations Assessment, Development and Evaluation instrument will be used to assess the quality of evidence in the NMA.3 PCI is the preferred method for acute myocardial infarction, and myocardial ischemia-reperfusion injury may occur after surgery, resulting in slow postoperative recovery of cardiac function in patients. In addition, PCI may promote the release of local inflammatory factors and vascular endothelial damage, leading to postoperative MACE. This study will discuss the efficacy and safety of TCMI in treating AMI from LVEF, hs-CRP, NT-proBNP, MACE, IL-6, and other aspects. As we all know, LVEF and NT-proBNP are important indicators to evaluate cardiac function, and the magnitude of NT-proBNP concentration can be applied to the early diagnosis of AMI and reflect the size of myocardial infarction area, suggesting cardiac function and quality of life of patients. hs-CRP is one of the factors that predict the risk of cardiovascular events, and its elevated level is positively correlated with cardiovascular risk. Studies have suggested that IL-6 level is closely related to long-term cardiovascular adverse events and mortality in patients with AMI.,37 TCMI has recently been shown to play an important role in antimyocardial ischemia-reperfusion injury, possibly associated with increased expression of Bcl-2 and decreased expression of Bax and Caspase 3, as well as Akt/eNOS signaling.,39As one of the common cardiovascular diseases, AMI has attracted extensive attention due to its rapid onset, rapid progression, high fatality rate, and poor prognosis. The incidence of AMI has been increasing year by year and is tending to be younger.This NMA was designed to evaluate the efficacy and safety of TCMI in the treatment of AMI, and to provide a basis evidence for the clinical practice of Traditional Chinese medicine. In our study, a comprehensive analysis was conducted on RCT of all TCMI treatments for AMI, including SMI, XMLI, YQFMI, SFI, DHI, HQI, SQFZI, etc, so as to synthesize these evidences to make a comprehensive ranking of available TCMI treatments for AMI. Furthermore, this study has some potential limitations. For example, most of the studies published in Chinese journals did not have a strictly randomized design. In addition, due to the limited ability of the author, only English literature and Chinese literature were searched, which may lead to the risk of bias.Conceptualization: Wei Zhao, Yawen Deng, Hengwen Chen.Data curation: Wei Zhao, Yawen Deng, Qingjuan Wu.Formal analysis: Wei Zhao, Hengwen ChenFunding acquisition: Jun Li, Hengwen Chen.Methodology: Wei Zhao, Yawen Deng, Qingjuan Wu.Project administration: Wei Zhao, Yawen Deng, Yuqing Tan.Writing – original draft: Wei Zhao, Yawen Deng.Writing – review & editing: Jun Li.


Background: The aim of this study is to report tumoral genetic mutations observed at high sequencing depth in a lung squamous cell carcinoma (SqCC) sample. We describe the findings and differences in genetic mutations that were studied by deep next-generation sequencing methods on the primary tumor and liver metastasis samples. In this report, we also discuss how these differences may be involved in determining the tumor progression leading to the metastasis stage.Methods: We followed one lung SqCC patient who underwent FDG-PET scan imaging, before and after three months of treatment. We sequenced 26 well-known cancer-related genes, at an average of ~6,000 × sequencing coverage, in two spatially distinct regions, one from a primary lung tumor metastasis and the other from a distal liver metastasis, which was present before the treatment.Results: A total of 3,922,196 read pairs were obtained across all two samples' sequenced locations. Merged mapped reads showed several variants, from which we selected 36 with high confidence call. While we found 83% of genetic concordance between the distal metastasis and primary tumor, six variants presented substantial discordance. In the liver metastasis sample, we observed three de novo genetic changes, two on the FGFR3 gene and one on the CDKN2A gene, and the frequency of one variant found on the FGFR2 gene has been increased. Two genetic variants in the HRAS gene, which were present initially in the primary tumor, have been completely lost in the liver tumor. The discordant variants have coding consequences as follows: FGFR3 , CDKN2A , and HRAS . The pathogenicity prediction scores for the acquired variants, assessed using several databases, reported these variants as pathogenic, with a gain of function for FGFR3 and a loss of function for CDKN2A. The patient follow-up using imaging with 18F-FDG PET/CT before and after four cycles of treatment shows discordant tumor progression in metastatic liver compared to primary lung tumor.Conclusions: Our results report the occurrence of several genetic changes between primary tumor and distant liver metastasis in lung SqCC, among which non-silent mutations may be associated with tumor evolution during metastasis. Lung cancer is the most common type of cancer and the leading cause of cancer-related deaths for both men and women in the world , 2. TherBecause of the heterogeneity of the tumor at genetic and molecular levels, treatment of SqCC continues to remain a challenge. There may be several reasons due to the existence of different genomic patterns in SqCC, and the genetic targets that have been found so far are probably not driver mutations and are therefore currently unactionable or “undruggable” mutations for this disease . SeveralThe spread of cancer cells from primary tumors to other vital organs is often associated with poor prognosis , 44. GivA 72-year-old African man was presented to our clinic in 2016, with a worsening dyspnea and chronic dry cough. Symptoms included fatigue and muscle atrophy, with pain in the chest and right rib cage. He suffered from cervical disc herniation and frequent reversible heart palpitation. The patient was a former smoker, consuming one pack per day for 16 years until he was 32 years old. In the previous two years, he had a significant weight loss of 20 kg. The physical exam shows a decrease in vesicular murmur and vocal vibrations of the right lung with no evidence of lymphadenopathy. The abdominal echography shows hepatomegaly. A suspicious mass for pulmonary neoplasm was found on chest X-ray. A CT scan detected a liver mass of 6 cm in the segment IV. Two biopsies were performed, one in the mediastinum (sample 69380-1-S7) and another in the liver (sample 69381-2-S8), and pathological examination confirmed the presence of squamous cell lung carcinoma, NSCLC at stage IVB (T4N2aM1c). The patient underwent FDG-PET scan imaging as part of the routine diagnosis and staging, before starting the chemotherapy treatment. Two hypermetabolic masses were localized; the maximum standardized uptake value (SUVmax) was assessed for these masses. On 26th December 2016, he received the first cure with paclitaxel–carboplatin. On 31st December 2016, he was admitted in the hospital for dysenteric syndrome and hemorrhoids and was treated with loperamide, diosmectite, metronidazole, lidocaine hydrochloride, diosmin, and hesperidin. On 19th January and 9th February 2017, he received the second and third cure with paclitaxel and carboplatin, respectively. Subsequently, on 13th March 2017, a second FDG-PET scan was performed to assess tumor progression. On 16th March 2017, he received the fourth cure with paclitaxel–carboplatin. A complete blood count was performed before each treatment, which was normal. A poor tolerance to chemotherapy was observed, as characterized with the following: nausea, diarrhea, loss of appetite, overall condition deterioration, and neurological disorders. Thus, the combination of immune checkpoint inhibitor and chemotherapy was not recommended and PD-L1 expression level was not investigated. Tyrosine kinase inhibitors (TKIs) were suggested instead. The patient refused to continue with the treatment. Nevertheless, he gave his consent to conducting the genetic analysis and for the publication of the present case report.https://www.agilent.com/cs/library/datasheets/public/TUMOR%20HOTSPOT%20MASTR%20PLUS%205991-8378ENN.pdf). This panel targets the following 26 genes: ALK, PTEN, PIK3R1, EGFR, NRAS, STK11, ERBB4, ERBB2, AKT1, HIST1H3B, HRAS, PDGFRA, MAP2K1, CDKN2A, IDH1, H3F3A, IDH2, BRAF, PIK3CA, KIT, CTNNB1, KRAS, FGFR3, MET, FGFR2, and DDR2.Microdissections from formalin-fixed paraffin-embedded (FFPE) specimens of these biopsies were performed, ensuring that a tissue area that contained a high percentage of tumor cells would be used for DNA extraction. gDNA was extracted using the QIAamp DNA Micro Kit (Qiagen), then the sequencing library was prepared and was submitted for sequencing on MiSeq (Illumina). We used an existing commercial gene panel test, the somatic Tumor Hotspot MASTR™ Plus with MID for sequencing library preparation (Multiplicom/Agilent). The assay developed for the identification of SNVs in 26 frequently mutated genes in solid tumors. This NGS assay was designed with the input from the French INCA centers  occurring in a high-complexity region, (2) having high, or higher than expected, variant fraction (germline), (3) having a sequencing coverage >50 ×, (4) having >50 supporting reads, and (5) being aligned inside the target region. The copy number variation analysis was not investigated in the present study. Variants were ranked by concordance and discordance between the two samples, and annotation information such as coding consequences was taken forward for further analysis. For each variant, we also reported the variant frequency, defined as variation percent =  × 100, where “altNum” is the number of reads containing the variant of interest and “depth” is the sum of number of reads containing the variant and the reads covering the reference allele. To obtain pathogenicity prediction scores, the target variants were submitted to the following databases made available by the Sophia Genetics DDM platform: COSMIC, Clinvar, OncoKB, cBioPortal, Varsome, Mutationassessor, Provean, FATHMM, MutationTaster, FATHMM-MK, MetaSVM, MetalR, LRT, SIFT, dbSNP, ClinVitae, and Provean. The protein structures encoded by each mutated gene were submitted to the SMART tool , to descTwo intense hypermetabolic masses were revealed by 18F-FDG PET/CT . They weThe mediastinal and liver samples were sequenced, resulting in 1,661,981 and 2,260,216 read pairs, respectively. Ninety percent of the total reads were above Q35, which is considered as a high threshold for read quality. The mapping of merged reads on the targets genes resulted in 1,628,242 reads (>97%) successfully mapped for the mediastinal sample and 2,134,774 reads (>94%) for the liver sample. In the present experiments, we achieved a high sequencing coverage of the target region. The mean target coverage in this study was around 6,000×. The target region coverage distribution for both samples is shown in In the lung sample, 12 out of the 26 genes sequenced presented with genetic changes, with 33 variants retained for high-confidence call and 28 discarded. In the liver sample, 13 genes presented with genetic changes, with 36 variants found and 24 discarded. The comparison between the distal metastasis and the primary tumor shows 30 shared variants and 6 distinct variants. When analyzing the variant frequency of the shared variants and using primary tumor data as baseline, we can observe several clusters .We found 30 variants shared between primary tumor and metastasis, distributed across 12 genes, and having a similar frequency between the two samples. Among these variants, 6 have genetic changes in the coding sequence of the following genes: ALK (3 variants), CDKN2A , 3.*9) on CDKN2A gene, and a missense c.746C>G (p.Ser249Cys) on the FGFR3 gene. The HRAS (p.Gln61Pro) variant is found in primary lung tumor with a frequency of 11.5% and completely absent in the liver metastasis. The CDKN2A (p.Leu16Profs*9) and FGFR3 (p.Ser249Cys) variants are present only in the liver metastasis at a frequency of 70.6 and 26.2%, respectively (www.ncbi.nlm.nih.gov/clinvar/), under the accession numbers SCV001250917 to SCV001250974.We found six discordant variants, covering the following genes: FGFR2, FGFR3, CDKN2A, and HRAS. Among these variants, only three variants had coding consequences as follows: a missense c.182A>C (p.Gln61Pro) on the HRAS gene, a frameshift c.47_50delTGGC  is located between two functional domains, in a relatively conserved position, and is predicted to have a damaging, disease-causing, and high effect on protein function . This is*9), is located at a relatively conserved position and predicted to have a high effect on protein function , is located in the Ras domain and has been found to be expressed in the upper aero-digestive tract and in thyroid cancers . This vaExtensive clinical research efforts and advancements that have been made since the human genome was sequenced have greatly increased our knowledge on cancer genetics. Several genetic alterations have been shown to initiate and sustain specific deregulated cellular signaling pathways involved in various tumors. In the present study, using one lung SqCC patient, we screened 26 genes that are well known as potential driver cancer genes, using next-generation sequencing. Though there are many known gene panels containing more genes, we limited our analysis to the current panel, to achieve high sequencing coverage. The panel that we used has been previously validated as clinically relevant in a wide variety of solid tumors. Moreover, this multiple-gene panel focuses on actionable hotspot mutations, which have therapeutic as well prognostic values for different solid tumors, enabling a personalized medicine approach.We performed in-depth investigation looking for SNVs and indels on the target genes. For each gene, both coding and noncoding regions were covered. We found that 83% of the retained variants are being shared by distant metastasis and primary tumor. Moreover, for each of these concordant variants, the variation frequency shows a strong concordance between the two tumor sites, with the presence of distinguished clusters, mainly containing both low- and high-frequency variants. Among the shared variants, we detected six non-silent SNVs, among which three variants at high frequency (36–99%) in the ALK gene, and at low frequency (1–6%): two in PIK3CA and one in CDKN2A. The variants CDKN2A (p.Leu65Pro) and ALK p.Ile1461Val) were found in another patient also with lung SqCC, with the same variant frequency (data not shown). Somatic alterations of ALK gene are reported relatively rarely in squamous cell carcinoma (SqCC) , but, on1Val wereIn the present report, six genetic variations affecting four genes  are shown to have a substantial discordance between the liver metastatic site and the primary tumor of the lung SqCC. Four genetic changes were gained on the genes FGFR2, FGFR3, and CDKN2A, and two genetic variants were lost on HRAS, in the metastatic tumor. Interestingly, after four cycles of treatment, a strong signal of resolution was detected at the liver metastasis through a significant sink over 60% of the tumor size. In contrast, the primary tumor showed a progression. These mixed outcomes after treatment may be related to the intra-tumor genetic heterogeneity of this lung SqCC patient. The discordant variants found in the current report have low frequency; it is possible that primary tumor cells with high-frequency genetic variants resist during the metastasis events, and a new clone harboring new low-frequency genetic variants emerges. These low-frequency variations might make metastatic tumor cells respond differently to chemotherapy treatment than the primary tumors.*9), and HRAS (p. Gln61Pro). None of these non-silent mutations were found in the study of De Bruin or Um , CDKN2A  is reported in somatic pancreas carcinoma and thymoma, also as a germline variant associated with hereditary cancer-predisposing syndrome. According to several databases, this variant is located at a relatively conserved position and is predicted to have a strong effect on protein function; the variant has a damaging, disease-causing effect on protein function (FGFR3 (fibroblast growth factor receptor 3) is a member of the fibroblast growth factor receptor family (FGFR). Alterations in the FGFR kinase family are common in lung SqCC and comprise the most frequently altered tyrosine kinase family , 51. Thefunction , 57. Thefunction . Until rfunction , to predfunction . In Um'sfunction .http://www.cruklungcentre.org/Research/TRACERx). One of the challenges faced in oncology is that the genotype does not correlate with the phenotype, for some patients. Consequently, this could result in significant inter-patient variability of drugs response, even for patients harboring the same molecular alteration. The molecular analysis of this report is restricted to SNVs and indels. For future study, it is important to expand the genetic analysis to other alterations and to assess their correlation with gene expression and protein levels.The metastatic event in lung cancer involves dissemination of cancer cells to anatomically distant organ sites, such as the liver and brain, and their subsequent adaptation to distant tissue microenvironments. Metastasis encompasses several biological processes, which consists of a complex series of well-organized steps: tumor cells exit their primary sites of growth by local invasion and intravasation, translocate systemically, survive in the circulation, arrest, affect the distant organ by extravasation, and adapt to survive and thrive in the initially healthy microenvironments of distant tissues , 44. MetIn this work, we shed light on cell molecular changes happening between the primary and metastasis tumors to the liver of lung SqCC, by targeting, at high sequencing depth, 26 well-known cancer-related genes. Our results show in lung SqCC the following: (1) the occurrence of genetic changes between primary tumor and distant metastasis to the liver, (2) the gain of function in the FGFR3 and probably the FGFR2 genes, and (3) the loss of function of CDKN2A. It would be interesting, for a more in-depth study, to investigate the prevalence of these variants in metastatic lung SqCC, in order to reveal the exact role of these variants in tumor evolution during each metastasis step and to assess the efficacy of treatments targeting these variants in the hope of opening opportunities for new efficient therapies for this disease.https://www.ncbi.nlm.nih.gov/clinvar?term=SUB7198155%5BSubmitter%20Batch%5D.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found below: Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. The patients/participants provided their written informed consent to participate in this study.Our major finding, we determined the genetic changes which occur between primary tumor and distal metastasis site in Lung SqCC.HM and AO designed and carried out the experiments. HM and RI analyzed the data. HM, VB, RI, and AO contributed to the interpretation of the results. HM wrote the paper with input from all authors. All authors discussed the results and commented on the manuscript.HM and RI are cofounders of the diagnostic device company OncoDiag SAS, Paris, France. HM is also a genetic counselor at the Al-Azhar Oncology Center, Rabat, Morocco. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The developed method was validated in terms of selectivity, linearity, accuracy, and precision.Bisphenol-A (BPA), bisphenol A glycerolate dimethacrylate (Bis-GMA), triethylene glycol dimethacrylate (TEGDMA), and urethane dimethacrylate (UDMA) are organic monomers that can be released from dental composites into the oral cavity. Over specific concentrations, they can act as endocrine disruptors or cause toxic effects. The purpose of this work is to develop and validate an analytical method to determine BPA, Bis-GMA, TEGDMA, and UDMA monomers released from synthetic dental resins in artificial saliva. The method was validated before its application to new hybrid ceramic materials used in computer-aided design and computer-aided manufacturing (CAD/CAM) restorations to determine the release of monomers in various time intervals , both in methanolic solutions, as well as in artificial saliva. Chromatographic analysis was performed isocratically on a Perfect Sil Target ODS-3 analytical column  with CH Dental amalgam has been in use for repairing dental cavities for almost 150 years. However, the Minamata convention for mercury release has affected the use of this traditional material due to concerns over its use and the potential release of mercury intraorally, and the environmental impact during its disposal [.Some of their disadvantages of dental fillings with amalgam are their metallic appearance, the high risk of tooth fracture, and the partial removal of tooth structure. There were some controversial studies about the possible mercury leakage, which can cause neuron and kidneys damage and other severe problems regarding human health. However, recent studies conclude that amalgam fillings are safe and should be placed in cases of uncooperative patients and special needs patients or patients with a high risk of caries . and environmental concern. Decreasing tolerance of exposure to other mercury compounds will certify that the use of mercury in dentistry remains an issue. Moreover, the Minamata conversion calls for a phase-down approach to dental amalgam  through greater emphasis and research into new dental materials as resin composites [. All the above have led to other dental materials, such as methacrylic resins and silicate cement. However, those materials were not as safe as they were expected to be, raising more research in order to establish the biocompatibility and safety of resin composites [However, it is clear from the research that mercury vapor during the removal of amalgam fillings is about ten times more toxic than lead on human neurons and with synergistic toxicity to other metals  and envimposites . All themposites ,5.Nowadays, a popular choice among dentists is dental polymer resin-based materials, which consist among others, of a polymeric organic matrix, inorganic fillers , and a coupling agent (γ-methacryloxypropyltrimethoxysilane), which is necessary for the chemical bondage between the two ,6. Resin, and better internal fit [ Therefore, resin-composite CAD/CAM materials are preferred by the clinicians for large MOD  cavities than amalgam and from ceramic CAD/CAM materials [. However, there is a limited number of studies that have evaluated the leaching of CAD/CAM materials [CAD/CAM technology is a rapid method for the fabrication of dental prosthetic restorations . Commonlrnal fit  comparedrnal fit . Therefoaterials . Howeveraterials . The orgaterials ,21. It iaterials ,23. Howeaterials . Bis-GMAaterials .The leaching of monomers such as UDMA, TEGDMA may have cytotoxic or allergic effects . In addiIn previous years, several liquid chromatographic methods have been developed for the determination of the monomers mentioned above. Some of them study the elution of monomers  from dental materials after their immersion in methanol or, most frequently, ethanol or deionized water, for a short or an extended period ,28,29,30The aim of this study is to develop and validate a reliable method to be used as the necessary analytical tool for dentistry to study the respective materials. Thus, a simple, accurate, and precise HPLC method for the simultaneous determination of BPA, Bis-GMA, TEGDMA, and UDMA in artificial saliva is presented herein.The CAD/CAM materials were selected according to the specific producer’s classification. Tetric CAD  has been characterized as a resin-composite CAD/CAM block incorporating an optimized mixture of cross-linked dimethacrylates and inorganic fillers; the material Vita Enamic  has been the representative of PICN (polymer-infiltrated ceramic network), which incorporates a porous sintered ceramic network filled with plastic. Furthermore, a newly introduced material is produced by filler press, and the monomer infiltration method (FPMI), the only representative of this kind of material, is the Katana Avencia block . The conventional resin composite Tetric  was selected due to the characterization as a classical resin composite restorative material .2, and KH2PO4 were supplied by Merck KGaA .Methanol, acetonitrile, and water of HPLC grade were obtained from PanReac AppliChem . BPA, TEGDMA, UDMA, and Bis-GMA were of analytical grade and were supplied by Sigma-Aldrich-LLC . Urea was purchased from Sigma-Aldrich . NaCl, CaCl2, and 0.26 g KH2PO4. Stock solutions of 100 ng/μL of standards were prepared by dissolving the appropriate amount of each monomer in artificial saliva and methanol.One liter of artificial saliva consisted of 1 L deionized water, 1 g urea, 0.552 g NaCl, 0.6 g CaCl3CN/H2O, 58/42% v/v as mobile phase. The samples were injected via a Rheodyne 7125 injection valve, with a loop of 20 μL volume . Detection was achieved at a wavelength of 215 nm and a sensitivity setting of 0.002 AUFS using an SSI 500 UV–vis detector .An LC–10 AD pump by Shimadzu  was used for the delivery of the mobile phase to the analytical column at a flow rate of 1.0 mL/min. Chromatographic separation was achieved isocratically in a PerfectSil Target ODS-3  analytical column  at room temperature, with CHStock solutions of 100 ng/μL of BPA, Bis-GMA, TEGDMA, and UDMA were prepared by dissolving the appropriate amount of each monomer in artificial saliva and methanol. Both methanolic and artificial saliva working standards were prepared in the range of 0.1–20 ng/μL for all monomers. Six calibration points were used at concentrations ranging from 0.5 to 20 ng/μL. Standard solutions were stored at 4 °C.Each CAD/CAM block material was sectioned into 10 × 10 × 2 mm square blocks were prepared using a 0.3-mm thick diamond-coated low-speed band saw  under sufficient water coolant. The total number of specimens for each of the CAD/CAM materials and the control material was eighty specimens (N = 80). In the center of each block, a small hole was made by using a 1.5-mm-diameter diamond rotary instrument. Ten specimens of each block were immersed in 8 mL of artificial saliva, and the other ten were immersed in deionized water. A silk thread passed through the hole in the middle of the block, and the specimens were immersed in the solution of each vessel. The glass vessels were airtight closed by using a screw cap. In this way, all surfaces of the specimens could be in direct contact with the medium, ensuring more representative results . All samThe number of specimens for all the groups was defined by statistical calculations. The statistical analyses of the pilot study were carried out using GPower 3.1.9.2 for Mac using the following statistical tests: ANOVA repeated measures within factors, a err prob = 0.05, power (1-b errprob) = 0.80, for three repetitions.3CN/H2O (58/42% v/v), within 23 min, on an HPLC system with a UV–vis detector set at 215 nm. Retention times were 5.5 min for BPA, 9 min for TEGDMA, 21.5 min for UDMA, and 22.5 min for Bis-GMA. Resolution factors were higher than 3.9, confirming the sufficiency of the separation.The separation of the monomers BPA, Bis-GMA, TEGDMA, and UDMA was achieved isocratically with a mobile phase consisting of CHTypical chromatograms of an artificial saliva blank solution, an artificial saliva standard solution, and a methanolic standard solution are given in The linearity was evaluated using mixtures of methanolic and artificial saliva standard solutions in different concentration levels within the working range. The upper limit of the linear range was 20 ng/μL for all monomers. Correlation coefficients ranged from 0.991 to 0.997 for artificial saliva solutions and from 0.994 to 0.999 for methanolic solutions. The limit of detection (LOD) and limit of quantification (LOQ) were calculated experimentally with the analysis of standard artificial saliva solutions of concentrations between 0.1 and 0.4 ng/μL. The LOQ was 0.2 ng/μL, and the LOD was 0.06 ng/μL. Calibration curves for each substance investigated were also constructed for standard artificial saliva solutions. The calibration and sensitivity data mentioned above are summarized in Accuracy and precision were examined at three concentration levels. Mean accuracy was expressed as the recovery (%) of the monomers from artificial saliva standard solutions and was found over 98% in all cases.n = 4) revealed RSD% levels below 15%, whereas on concentrations a lot higher than the LOQ RSD% was found under 1%. Inter-day precision measured over a period of five days revealed RSD% values higher than those of the intra-day measurements, but still lower than 15%. The results are summarized in Intra-day precision  14 days of immersion in artificial saliva is illustrated in A simple, accurate, and precise method for the simultaneous determination of BPA, Bis-GMA, UDMA, and TEGDMA from resin-based dental composites in artificial saliva is described herein. For the validation of the developed method, linearity, sensitivity, accuracy, and precision were evaluated. Repeatability and precision, expressed by RSD values, were lower than 15% in most cases. The relative errors ranged from 98% to 106% for artificial saliva standards.To the authors’ knowledge, there are few articles in the literature to determine the leaching monomers from CAD/CAM materials ,35. HoweThe lower monomer leaching from CAD/CAM materials, compared to conventional resin-based materials, could be attributed to the different methods of fabrication and the difference of the organic matrix of the materials. The composition of the CAD/CAM materials clearly indicates that the manufacturers have interpolated UDMA and TEGDMA as the preeminent monomer . The advIn terms of application to real samples, there was a decreasing pattern for the monomers that were eluted, considering the time intervals for the CAD/CAM materials. This elution stopped completely 28 days after the first immersion of the CAD/CAM materials in artificial saliva or distilled water for all monomers. The most eluted monomer in those materials seems to be TEGDMA. Due to its low molecular mass and its high mobility and activity, its elution from the organic matrix is more likely to happen, compared to the other, less flexible monomers. On the contrary, there was a constant release of all monomers for the conventional resin composite, even 28 days after its first immersion in artificial saliva or distilled water. The above can be attributed to the different methods of fabrication of resin-composite CAD/CAM materials, which consists of high-temperature or/and high-pressure polymerization.The herein developed HPLC method is fully validated and proved to be suitable for its purpose as it is very accurate and precise for the newly introduced resin-composite CAD/CAM materials. It requires minimal sample preparation, and the instrumentation involved are inexpensive and straightforward.


Sensitivity, specificity, positive and negative predictive values, and diagnostic accuracy for GCs were 73%, 90%, 89%, 75%, and 81%, respectively. On multivariate analysis, lesion detectability on plain CT was independently associated with GCs . Histopathologically, GChPs consisted of adipose tissue. Although larger vessel areas in GCs than in GChPs was not significant , cell densities were significantly greater in GCs . Among GPs larger than 10 mm, plain CT could contribute to differentiating GCs from GChPs.An appropriate diagnosis is required to avoid unnecessary surgery for gallbladder cholesterol polyps (GChPs) and to appropriately treat pedunculated gallbladder carcinomas (GCs). Generally, polyps >10 mm are regarded as surgical candidates. We retrospectively evaluated plain and contrast-enhanced (CE) computed tomography (CT) findings and histopathological features of 11 early GCs and 10 GChPs sized 10–30 mm to differentiate between GC and GChP >10 mm and determine their histopathological background. Patient characteristics, including polyp size, did not significantly differ between groups. All GCs and GChPs were detected on CE-CT; GCs were detected more often than GChPs on plain CT (73% vs. 9%;  A gallbladder polyp (GP) is any type of elevated lesion of the gallbladder ,2. GPs aTo differentiate between benign and malignant GPs, various methods have been reported, including high-resolution abdominal ultrasonography (AUS) , contrasThe CT number is used to reveal the signal intensity of each part of the image, which is estimated from the difference in the absorption coefficient between water and the object ,15. The Generally, GPs with a diameter larger than 10 mm are considered to require treatment ,17,18. HWe retrospectively evaluated the findings on plain CT and contrast-enhanced CT (CE-CT), and the histopathological features of GCs and GChPs. We also compared the detectability on plain CT and the histopathological characteristics associated with their detectability.We investigated patients who underwent cholecystectomy at Shizuoka Cancer Center between January 2005 and December 2018. During this period 114 GCs were resected, and all were included in the study. There were 12 early GCs with polypoid growth with a diameter of 10 to 30 mm. One patient without CE-CT due to renal dysfunction was excluded, and 11 out of the 12 early GCs were analyzed in our study. As a control group, 10 consecutive GChPs with a diameter of 10 to 30 mm resected in the same time period were analyzed. The size of the GP was measured on CE-CT. If the patient did not undergo CE-CT or if the GP could not be detected by CE-CT, polyp size was measured on abdominal ultrasound. The study was conducted according to the guidelines of the Declaration of Helsinki, and approved by the Institutional Review Board of Shizuoka Cancer Center .CT scans were performed using multidetector-row CT with a slice thickness ≤ 5 mm, and the features of CE-CT were evaluated during the late phase (180 s after injection of the contrast medium). All patients except for one GChP patient received CE-CT within 3 months before surgery. Two gastroenterologists  interpreted the detectability of lesions using CT, without information regarding the diagnosis. If both physicians could detect the lesion, the detectability was considered positive . The repThe CT number of each lesion was measured twice, and the mean value was calculated. If the lesion was not detected, then the CT number was measured at the part that was identical to that of the lesion detected on CE-CT. On CE-CT, the CT number of the lesion was measured at the venous phase. Simultaneously, the CT number of bile was measured around the lesion.Surgically resected specimens were examined according to the General Rules for Surgical and Pathological Studies on Cancer of the Biliary Tract of the Japanese Society of Biliary Surgery . The nump < 0.05 was considered statistically significant. All analyses were performed using R version 3.4.1 .Continuous variables were analyzed using the non-parametric Mann–Whitney U test, and categorical and binary variables were analyzed using Fisher’s exact test due to the relatively small number of cases. All statistical tests were two-tailed and Age, sex, polyp size, carcinoembryonic antigen value, and CA19-9 value did not significantly differ between the GC and GChP groups . There wp < 0.01)  . Three cp = 0.045)  .2 vs. 0.0080/μm2; p < 0.01) , the difference did not reach significance  (p = 0.04)  . Althougificance . Cell de < 0.01) . The cel = 0.04)  and the The Clinical Practice Guideline for the Management of Biliary Tract Cancers  indicateMany reports have described that a GP larger than 10 mm could be a GC ,17,18, aVarious reports have indicated the differential diagnosis of GPs ,13,26,27The number and area of the nuclei were higher in GCs than in GChPs. This suggests that the cell densities are greater for GCs than those for GChPs, thereby contributing to the larger CT number for GCs. The CT number is calculated by the difference in the absorption coefficient of the X-ray between water and tissue; therefore, the density of the tissue is related to the CT number, and higher-density tissue has a higher CT number .Moreover, GChPs have abundant lipids in the mucosa propria, and the lipid content generally lowers the CT number of the tissue. These things are associated with the result that GChP could be rarely recognized.The vessel areas of each tissue field were larger for GCs than for GChPs. The CT attenuation of blood reflects the concentration proteins including hemoglobin which has a high density. However, the difference between GCs and GChPs was not statistically significant.If a GP larger than 10 mm is recognized on plain CT, then GC is suspected, and open surgery is recommended . A GP noOur study was limited by its retrospective, single-center design. Additionally, it was limited by the small number of cases. A further prospective confirmatory study is necessary to clarify our findings.Among GPs larger than 10 mm, plain CT could contribute to differentiating GCs from GChPs, i.e., GCs were detected significantly more often on plain CT than were GChPs, as GChPs were scarcely recognized. The ability to differentiate between the two depended on the lipid content and cell density of the lesion.


This study aimed to evaluate the potential relationship between oxidative stress, dietary intake, and serum levels of antioxidants in patients with a complete hydatidiform mole (CHM) compared with controls.®, Siemens).This was an observational cross-sectional study conducted in Rio de Janeiro, Brazil. A total of 140 women were enrolled in this study and divided into four groups: 43 patients with CHM, 33 women who had had an abortion, 32 healthy pregnant women, and 32 healthy non-pregnant women. All participants underwent blood sampling, assessment using a semiquantitative food frequency questionnaire, and anthropometric measurement. Blood samples were collected after overnight fasting (10-12 h). Vitamin levels  were determined by ultra-performance liquid chromatography, and gamma-glutamyl transferase levels were assessed using an automated quantitative analysis system (DimensionAlthough all groups showed sufficient serum vitamin A and E levels, the participants had inadequate dietary intake of these vitamins. Conversely, all groups had an insufficient serum level of vitamin C, despite adequate intake. The gamma-glutamyl transferase values did not differ significantly among the groups. However, elevated serum levels of this enzyme were observed in several patients.All groups exhibited high levels of oxidative stress, as evaluated by gamma-glutamyl transferase levels, and had inadequate intake of antioxidant vitamins. Therefore, the high exposure to oxidative stress found in our study, even in healthy pregnant and non-pregnant women, may increase the incidence of CHM in this region. Hydatidiform mole (HM) is a benign disorder resulting from an abnormal fertilization. It is characterized by atypical swelling of the placental villi and intense trophoblastic hyperplasia . It is tIn Brazil, the estimated incidence of HM is 1 per 200-400 pregnancies, which is 5-10 times higher than that in Europe and the United States . This discrepancy in the incidence of HM worldwide could be attributed to methodological differences or the lack of centralized databases, or it could be linked to genetic, socioeconomic, and nutritional factors . AlthougThe etiology of HM is not yet completely understood . HoweverThe main components of human non-enzymatic antioxidant defense mechanism encompass dietary vitamins A, C, and E and glutathione (GSH). Nutritional deficiencies in these elements are potential risk factors for CHM ,4. DecreVitamins A and E are considered important dietary antioxidants because of their ability to reduce oxidative activity in lipophilic environments. Vitamin A is well established as an efficient scavenger of free radicals , while vGSH plays a central role in the intracellular non-enzymatic antioxidant system. Besides neutralizing free radicals, GSH is essential in the regeneration of vitamin C . The metSupplementation with antioxidant vitamins, like A, C, and E, has been suggested as a strategy for the prevention of CHM and its progression to GTN ,18. HoweThis was an observational cross-sectional study conducted in two Brazilian GTD reference centers (RCs) located in the state of Rio de Janeiro at the Antonio Pedro University Hospital  and the Maternity School (UFRJ). Both RCs are located in public hospitals that are part of the Brazilian Unified Health System.The study was approved by the Human Research Ethics Committees of the Medical School, UFF (CAAE: 37660714.7.0000.5243) and the Maternity School, UFRJ (CAAE: 38329214.0.0000.5275). All procedures performed were in accordance with the ethical standards of the institutional and/or national research committee as well as the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all individual participants included in the study.We enrolled 140 women in this study, of whom 43 women had a CHM diagnosis confirmed by histopathological examination (CHM group), 33 women had histopathological examination findings indicative of spontaneous non-molar abortion , 32 were healthy women at 20 weeks of gestation with a single viable fetus , and 32 were healthy non-pregnant women .Every pregnant woman visiting the RCs who was suspected of having a molar pregnancy and had undergone molar evacuation was invited to participate in the study when they arrived for their first medical appointment after uterine evacuation. After histopathological diagnosis, these patients were allocated to two goups according to their diagnosis: complete hydatidiform mole (CHM) group and non-molar abortion (Ab) group. The patients in the other two groups (HP and NP groups) were recruited in the non-GTD clinics and paired by age with those in the CHM group. For the healthy pregnant group, only participants with a gestational age of up to 20 weeks were recruited.®, Siemens).As most cases of uterine evacuation are not elective, the team decided to conduct the recruitment during the first visit to the RC after uterine evacuation. This ensured an overnight fasting period (10 to 12 hours) before venous blood collection, as well as guaranteed a regular dietary intake the day before blood collection. Vitamin levels were determined by ultra-performance liquid chromatography. GGT, aspartate aminotransferase, alanine aminotransferase, and bilirubin were assessed using an automated quantitative analysis system ], according to Lohman et al. , height (m), and body mass index .For the evaluation of vitamin intake, a semiquantitative food frequency questionnaire (SFFQ) was used. The food list in the SFFQ was determined based on the foods mostly consumed by the Brazilian population . The freMeasures . The resSince the recommended dietary allowance (RDA) for vitamins varies according to age and is increased during gestation, the estimated intake (mg/day) has been converted to intake adequacy (%). The RDAs used to calculate the intake adequacy were as follows: vitamin A=700 µg/day for non-pregnant women, 750 and 770 µg/day of vitamin A for pregnant women , vitamin C=65 mg/day and 75 mg/day for non-pregnant women ; 80 mg/day and 85 for pregnant women , and vitamin E=15 mg/day for all participants.t-test or a Mann-Whitney test with Bonferroni correction. The frequencies were compared using the Fisher's exact test and Pearson’s chi-square test as appropriate. All statistical analyses were performed using R version 3.2.2, and p-values of <0.05 were considered significant.Given that the previous literature has mostly focused on molar pregnancy and low vitamin A intake, the study was powered to see the difference of at least 0.1 mg/L below the lower limit of the vitamin A reference range of 0.3 mg/L. At least 24 women were needed in each group to detect a difference between 0.3±0.1 mg/L and 0.2±0.1 mg/L at 80% power at an alpha level of 0.01. Data were expressed as medians, first and third quartile, or frequencies and percentages, as appropriate. Continuous variables were evaluated for normality using the Shapiro-Wilk test. For variables with normal distribution, the groups were compared using analysis of variance; for variables with non-normal distribution, Kruskal-Wallis test was used followed by a non-parametric Student’s The demographic and laboratory characteristics of the studied population are shown in p=0.004). On the contrary, the vitamin A serum levels in the Ab and NP groups were higher than those in the CHM group . Although significant differences were observed among the groups, all groups showed normal median vitamin A serum levels, with serum concentrations within the reference interval (0.3-0.7 mg/dL). Similarly, with regard to intake adequacy, all groups had a median adequate intake of this vitamin, with no significant difference between the groups (p=0.215). However, the prevalence rates of vitamin A intake inadequacy were 48.8% in the CHM group, 37.5% in the Ab group, 40.6% in the HP group, and 46.8% in the NP group.The median serum levels and intake adequacy of antioxidant vitamins are described in p=0.624). Conversely, all groups showed a median vitamin C intake over 100% with no significant difference between groups (p=0.958). However, the prevalence rates of inadequate vitamin C intake were 39.5%, 42.4%, 28.1%, and 34.3%, respectively, in the CHM, Ab, HP, and NP groups.As regards vitamin C, the serum levels were compatible with the classification of deficiency, which means none of the four studied groups achieved the lower limit of the normal range (0.5 mg/dL). Circulating vitamin C deficiency was observed in 71% of the studied population and did not differ significantly among the groups (p=0.675). However, in terms of the daily vitamin E intake, all groups had a median vitamin E intake adequacy lower than 100%, with no significant difference among the groups (p=0.052). Moreover, most patients in all groups did not take enough vitamin E, demonstrating prevalence rates of inadequate dietary intake of 55.8% in the CHM group, 72.7% in the Ab group, 53.1% in the HP group, and 75% in the NP group.With regard to vitamin E, all participants showed normal serum levels, with serum concentrations above the reference value for this vitamin (12 μmol/L). No statistically significant difference was found among the four groups (p=0.559) and Q4 (p=0.084), presenting similar exposure to OS.When we analyzed the serum GGT levels as an OS marker, the quartiles and median of the studied population were as follows: median: 24 U/L, first quartile (Q1): ≤18 U/L, second quartile (Q2): >18 and ≤24 U/L, third quartile (Q3): >24 and ≤33 U/L, and fourth quartile (Q4): >33 U/L). The population distribution, in percentage, of each studied group distributed into GGT quartiles, is shown in Pregnancy is characterized by a physiological increase in free radical production and, consequently, increased recruitment of antioxidant defenses. Thus, an adequate supply of antioxidant vitamins in this period is essential to ensure the maintenance of oxidative balance. Moreover, OS plays an important role in the pathogenesis of CHM ,7,8, andOur results on serum vitamin A disagree with those reported by Andrijono et al.  and KoluVitamin C, along with vitamin E, comprises the first line of defense and is the most important of the non-enzymatic diet antioxidants ,17,19. TWe found a high prevalence of serum vitamin C deficiency in more than 70% of the participants. Oliveira et al. reported 30.8% C-hypovitaminosis in pregnant women from southeastern Brazil, the same region where this study was conducted . The higWith regard to the serum levels of vitamin E, our results disagree with those of Kolusari et al., who reported a decrease in serum levels of vitamin E among patients with CHM . We did Vitamin E sufficiency is described as serum concentrations higher than 12 μmol/L; however, many studies have already indicated that optimal levels, greater than 30 μmol/L, have benefits in the prevention of several types of cancers ,30. In oHowever, despite the serum sufficiency and similar to what was described by Péter et al. regarding vitamin ingestion,  we obserOS has already been associated with the pathogenesis of CHM since higher concentrations of its markers and lower levels of antioxidants have previously been observed in women with this disease -8. In adA novel aspect of the present study is the finding that GGT is a systemic and early OS marker among patients with CHM. Our findings corroborate with those of other studies reporting that elevated levels of GGT are associated with inflammation and subclinical OS in a healthy population ,19. ThusThis study has some limitations. During the gestational period, the woman undergoes several physiological changes to ensure the development of the fetus, and these metabolic changes are reflected in laboratory tests at the time of blood collection. In contrast, CHM and AB groups blood samples collection were performed after uterine evacuation. This condition may affect the comparison of samples between the groups. Although the samples included in this study were statistically adequate for analyzing vitamin A, it would be appropriate to carry out a more extensive case series evaluation not only to study the parameters analyzed with a higher power but also to evaluate the relationship between nutrition, other vitamins (such as C and E), and the progression of molar pregnancy to GTN. Moreover, patients in all four groups in our study were managed in two public, free-care hospitals, and their nutritional status may differ from those in studies conducted in populations with greater resources ,10. The While it is believed that OS and nutrition are closely related to the pathogenesis of CHM, other studies are necessary to elucidate this relationship. In our study, nutritional deficiency has been observed equally in women with CHM, as among those with aAb, HP women, and NP healthy women. Therefore, until more extensive results are published, it may not be appropriate to advocate nutritional supplementation strategy specifically as a way to prevent CHM. Given the frequent nutritional deficiencies in the Brazilian population, a healthy diet, especially in those that intend to become pregnant, should be recommended and supported for all women.Ramos CAB, Ferraz L, Braga A and Lopes PF conceived and designed the study and were responsible for fulfilling the ethical requirements during the design and execution of the study. Ferraz L was responsible for all nutritional assessments. Ramos CAB and Ferraz L were responsible for conducting the biochemical evaluation, audited by Lopes PF. Velarde LGC was responsible for the sample size calculation and statistical analysis. All authors contributed to the data analysis, interpretation, and manuscript writing and final approval.


Glioma is one of the leading causes of death from cancer, and autophagy-related genes (ARGs) play an important role in glioma occurrence, progression, and treatment. In this study, the gene expression profiles and clinical data of glioma patients were obtained from The Cancer Genome Atlas (TCGA) and the Chinese Glioma Genome Atlas (CGGA), respectively. ARGs were obtained from the Human Autophagy Database. We analyzed the expression of the ARGs in glioma and found that 73 ARGs were differentially expressed in tumor and normal tissues. Univariate Cox regression analysis was used to identify prognostic differentially expressed ARGs (PDEARGs). Least absolute shrinkage and selection operator (LASSO) and multivariate Cox regression analyses were performed on the PDEARGs to determine the risk genes; and BRIC5, NFE2L2, GABARAP, IKBKE, BID, MAPK3, FKBP1B, MAPK8IP1, PRKCQ, CX3CL1, NPC1, HSP90AB1, DAPK2, SUPT20H, and PTEN were selected to establish a prognostic risk score model for TCGA and CGGA cohorts. This model accurately stratified patients with different survival outcomes, and the autophagy-related signature was also appraised as being an independent prognostic factor. We also constructed a prognostic nomogram using risk score, age, gender, WHO grade, isocitrate dehydrogenase (IDH) mutation status, and 1p/19q co-deletion status; and the calibration plots showed excellent prognostic performance. Finally, Pearson correlation analysis suggested that the ARG signature also played an essential role in the tumor immune microenvironment. In summary, we constructed and verified a novel autophagy-related signature that was tightly associated with the tumor immune microenvironment and could serve as an independent prognostic biomarker in gliomas. Glioma is one of the most common primary brain tumors in adults, accounting for about 15% of all brain tumors and more than 48% of malignant brain tumors , 3. For To illustrate the importance of autophagy, there is increasing evidence of the role of autophagy-related genes (ARGs) in the development of cancer. High expression of Atg10 is associated with poor prognosis for lung cancer . AutophaAlthough some previous studies have explored the relationship between ARGs and prognosis in glioma patients, those studies have focused on the function of a single gene. Very few studies have used gene expression profiles to examine the relationship between multiple ARGs and glioma prognosis. Therefore, in this study, we develop a reliable prognostic model based on ARGs and explore the clinical application of the ARG signatures in glioma patients.1 and used as a training cohort. From the Chinese Glioma Genome Atlas (CGGA) dataset2, we collected the RNA-Seq data of 1,018 samples as a validation cohort (3 (The RNA-Seq and somatic mutation data of 703 patients were downloaded from The Cancer Genome Atlas (TCGA) data portal 1 and usen cohort , 16. We ohort  in glioma and normal neural tissue samples . A WilcoP-value of <0.05  by using the “survival” package with a of <0.05 . Second,of <0.05 . Third, of <0.05 . The folCoef is the multivariate Cox regression analysis coefficient of the ARGs and X is the relative expression level of each ARG. The patients were divided into high-risk and low-risk groups, with the median risk score used as the cutoff value and high-risk scores suggesting that the prognosis of the patients was poor.where t-tests were used to assess differences between the variables. Kaplan–Meier curve and log-rank tests were used for survival data analysis, and univariate Cox regression analysis was used for survival factor analysis. Multivariate Cox regression analysis was used to identify independent prognostic factors. The accuracy of the prognostic model was assessed by time-dependent receiver operating characteristic (ROC) analysis. A P-value of <0.05 was considered statistically significant.R software (3.6.1) was used for all statistical analyses. Pearson correlation coefficients were used to evaluate the rank correlations among the variables. Independent n = 698) and brain tissues (n = 5) in TCGA cohort were examined, and these values were compared using the Wilcoxon signed-rank test. This analysis revealed 15,001 DEGs in glioma tissues compared with normal brain tissues . Then, we obtained 73 DEARGs in glioma tissues compared with normal brain tissues after taking the intersection of DEGs and ARGs .To identify the PDEARGs, we performed a univariate Cox regression analysis of each DEARG expression in TCGA cohort. A total of 62 DEARGs were significantly correlated with OS in glioma patients . MAPK3 and CX3CL1 were strongly positive; NFE2L2, HSP90AB1, and SUPT20H were moderately positive; and FKBP1B, BIRC5, NPC1, IKBKE, BID, and PTEN were weakly positive in glioma tissue relative to their expression levels in normal tissue (P < 0.05) had better prognosis (P < 0.05) had better prognosis  and a low-risk group (n = 333), according to the median risk score. We used a Kaplan–Meier curve based on the log-rank test to identify the difference in prognosis between the high-risk group and the low-risk group. The prognosis of patients in the high-risk group was worse than that in the low-risk group (P < 0.05) (Patients in TCGA cohort were divided into a high-risk group ( < 0.05) .We then used time-dependent ROC curves to confirm the accuracy of the model’s predictions for 1–10 years. The ROC (AUC) of the prediction model was 0.906 over 1 year, 0.921 over 2 years, 0.924 over 3 years, 0.89 over 4 years, 0.874 over 5 years, 0.882 over 6 years, 0.855 over 7 years, 0.809 over 8 years, 0.822 over 9 years, and 0.836 over 10 years in TCGA training cohort , all of We also generated heatmap to describe the expression patterns of risk genes in both the high-risk and low-risk groups . In patiP < 0.05)  , with th < 0.05) , which w < 0.05) . The ris < 0.05) . These rIn addition, when the patients were stratified according to different clinicopathologic parameters, the autophagy signature remained a significant prognostic factor in both TCGA cohort  and the P < 0.05) (P < 0.05), while WHO grade, 1p/19q co-deletion status, and risk score were independently associated with OS in the CGGA cohort (P < 0.05) ( < 0.05) ; only ge < 0.05) . These rTo better predict the prognosis of patients with glioma at 1–10 years, we constructed a new nomogram . In FiguBy analyzing somatic mutation data from TCGA dataset, we explored differences in genomic changes between the high-risk and low-risk groups, as some genomic changes have a negative impact on glioma survival . By compP < 0.05). These results demonstrate that the dysregulation of autophagy-related risk gene expression is associated with the development of glioma.To examine the performance of our model in predicting the progression of glioma, we analyzed the relationships between the clinical variables in the two cohorts and the risk genes. We found that the expressions of BRIC5, FKBP1B, IKBKE, and NFE2L2 were significantly greater in the 1p/19q non-co-deletion samples, while the expressions of BID, CX3CL1, DAPK2, GABARAP, HSP90AB1, MAPK3, MAPK8IP1, and SUPT20H were significantly lower in those samples in both cohorts ; the expTo determine whether the model could reflect the status of the tumor immune microenvironment in glioma patients, we analyzed the correlation between the risk scores and immune cell infiltration by using Pearson correlation analysis in both TCGA and CGGA cohorts. We found that the risk scores were correlated positively with the levels of CD8+ T cells; gamma delta T cells; M0, M1, and M2 macrophages; and activated dendritic cells. Meanwhile, they were negatively correlated with the levels of naive B cells, naive CD4+ T cells, follicular helper T cells, activated NK cells, monocytes, and resting mast cells, suggesting that the immune-related gene signature could indicate the level of infiltrating immune cells in gliomas to a certain extent . We alsoGlioma is a progressive disease that requires an effective prognostic indicator for diagnosis and treatment. There is increasing evidence that autophagy plays a key role in the occurrence, progression, and therapeutic resistance of glioma , 8, 25. In this study, we identified prognostic ARGs and used them to construct a reliable model for predicting the OS of patients with glioma. First, we analyzed the expression of 232 ARGs in glioma and found that the expression of 62 DEARGs was related to OS. These results suggest that ARGs are an important prognostic factor for glioma patients.Next, we evaluated these PDEARGs in the stratification of patient outcomes. Through a combination of LASSO regression and Cox regression analysis, we identified 15 PDEARGs of interest and used them to construct a Cox regression risk model. We also analyzed and verified the reliability and stability of the model. The results showed that the model can accurately discriminate patients with different survival outcomes. Univariate and multivariate Cox regression analyses indicated that the prognostic risk model could independently predict the prognosis of patients with glioma. In addition, nomogram analysis indicated that the accuracy of the model in predicting glioma prognosis could be improved in combination with other clinical features.Nomograms have been widely used in clinical work for visual presentation , 30. In The relationship of risk genes with clinical variables  was also analyzed. We found that risk genes were associated with the progression of gliomas. Our model therefore has high clinical application value in predicting the development of glioma.in vitro and in vivo by impacting the Hippo pathway , and Couldwell et al., found a correlation between PKC activity and glioma cell proliferation, as glioma cells with a fast development speed have high PCK activity . MAPK3 iin vitro .Previous studies have demonstrated that immune infiltration plays a crucial role in determining therapeutic effects in glioma and the prognosis of patients with glioma , 50. ThePrevious studies have also demonstrated that the expressions of immune checkpoint and HLA complex are an important factor in determining the response to treatment and the prognosis of patients with glioma –55. Lemkin vivo and in vitro experimental studies.There are two main limitations of the present study. Firstly, clinical information downloaded from TCGA and CGGA databases is limited and incomplete, and secondly, the identified mechanism of ARGs affecting glioma development requires further In summary, we used 15 ARGs to construct a prognostic risk model that accurately predicted the prognosis of glioma patients. The risk score generated by this model can be used as an independent prognostic indicator to discriminate patients with different survival outcomes. The model can also predict the immune microenvironment of glioma to a certain extent. However, further studies are needed using a prospective, large-scale, multicenter clinical cohort to validate the prognostic model.https://portal.gdc.cancer.gov/, http://www.cgga.org.cn/.Publicly available datasets were analyzed in this study. This data can be found here: CZ, KY, and RL conceived and designed the study, edited the manuscript, and acted as overall guarantors. YX and RL carried out the analysis procedure, contributed to the study design and interpretation of the results, and contributed to the drafting of the manuscript. XL, ND, DW, and LH contributed to the preparation of the dataset and drafts of the manuscript. All the authors reviewed the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


We report a case of cytomegalovirus- (CMV-) induced buccal ulcer in a patient with nephrotic syndrome. An 82-year-old man with membranous nephropathy was on immunosuppressive therapy presented with an ulcer in the oral cavity and was hospitalized. Intraoral examination revealed an inflamed and painful ulcer on the left buccal mucosa. Blood test results showed CMV positivity, and histopathological examination confirmed the diagnosis. Anti-CMV therapy (ganciclovir) was initiated from the third day of hospitalization. However, he developed dyspnea on the 14th day. Computed tomography images of the chest revealed the presence of ground-glass opacities, and noninvasive positive pressure ventilation was initiated under the provisional diagnosis of pneumocystis pneumonia caused by ganciclovir-associated myelosuppression and/or steroid-induced immunocompromised state. The patient died of pneumocystis pneumonia on the 21st day. The patient had received immunosuppressive therapy for renal dysfunction. Immunocompromised patients with CMV infection should be treated with caution, as drugs for CMV may themselves cause myelosuppression, deteriorating the prognosis of the patient. Cytomegalovirus (CMV) is a genus of viruses of the order Herpesvirales, which is estimated to infect approximately 90% of adults at some point in their lives. CMV infection is usually subclinical and can persist in this manner for the entire life span of an infected individual –3. It isTo date, however, few reports have documented the manifestations of CMV infection in the oral cavity –12. HereAn 82-year-old man was referred to our hospital from another clinic on June 11, 2019; at that time, he had low total protein (4.8 g/dL)/albumin levels (1.6 g/dL) and high protein-creatinine ratio (7.3) and was thus admitted to our department of nephrology. His medical history revealed frequent tonsillitis in childhood and two episodes of hemorrhagic gastric ulcers at the age of 76 years. He was not positive for HIV, and no family history of renal failure or history of consanguineous marriage was reported.He was diagnosed with membranous nephropathy based on the findings of a renal biopsy and was treated according to the standard treatment protocol , which iOn July 26, he started experiencing pain and difficulty in chewing due to pain in the entire oral cavity, and on August 5, he was admitted to our hospital on an emergency basis due to the presence of a severely painful lesion in the mouth.4/μL; and leukocyte count, 35.9 × 102/mm3. The results of further antibody tests revealed that he was positive for CMVpp65 antigens  546/434. His β-D glucan concentration was 40.2 pg/mL, suggesting a strong possibility of CMV infection.At the time of admission on August 5, 2019, his height and weight were 160 cm and 62.9 kg, respectively. Results of blood tests revealed the following: albumin concentration, 1.3 g/dL; C-reactive protein concentration, 1.39 mg/dL; total protein concentration, 4.1 g/dL; platelet count, 14.7 × 10On August 6, 2019, the patient presented with a painful intraoral ulcer; intraoral examination revealed ulcers on the left buccal mucosa and generalized inflammation of the gingiva. Moreover, erosion was observed on the left side of the tongue . No ulceA clinical diagnosis of ulcer on the left buccal mucosa was made, and the attending physician requested a biopsy of the ulcer for a definitive diagnosis. An incisional biopsy was performed, and the specimen was sent for a pathological examination.Histopathological findings revealed a prominent ulcerative lesion containing numerous CMV-positive vascular endothelial cells scattered at the fundus of the ulcer Figures  and 3. BThe pain persisted even after admission, and the patient was prescribed an azulene sodium sulfonate gargle containing 4% xylocaine. A 10-day course of anti-CMV medication (ganciclovir) was initiated on August 7, 2019.2/mm3, respectively  was 25.80 mL/min and blood urea nitrogen (BUN) was 60 mg/dL, while on August 19, eGFR decreased to 19.42 mL/min and BUN increased to 73 mg/dL. A rapid decline in his renal function was evident. Sulfamethoxazole-trimethoprim (10 mL/day) was initiated from August 20. However, the patient died due to respiratory failure on August 25, 2019.Before his death, the patient provided written informed consent for the publication of his images and case details. The requirement for ethical approval was waived by the appropriate institutional review board.The potential routes of CMV infection include vertical transmission via the placenta, birth canal, and/or breast milk; horizontal transmission via the saliva, urine, semen, blood, and/or cervical and vaginal discharge; or organ transplantation. In many cases, CMV infection occurs during the perinatal period or infancy, and many infected individuals are asymptomatic . HoweverA few reports of intraoral CMV infection , 14 in tIn contrast, few reports have described oral ulcers caused by CMV in patients with adult nephrotic syndrome, presenting with fatigue, edema, and proteinuria , rather A diagnosis of CMV infection can be confirmed by virus isolation; the presence of antigenemia, which detects polymorphonuclear leukocytes that are positive for the viral antigen pp65 , 18; polIn the specimen obtained from our patient, the nuclei and cytoplasm of hematoxylin and eosin-stained cells were positive for the viral antigens, and positive cells were also observed during the immunohistopathological examination. Particularly, vascular endothelial cells show high affinity for CMV and harbor the virus . The infIn principle, antiviral medications such as ganciclovir are indicated for CMV infections  , 19, 20.The patient presented with renal dysfunction and developed pneumocystis pneumonia after the onset of the clinical CMV infection. Pneumocystis pneumonia is a serious manifestation of an opportunistic pathogen such as CMV, and previous reports have documented concurrent CMV infection and pneumocystis pneumonia in patients with HIV .The dose and duration of ganciclovir therapy with respect to immune dysfunction were appropriate. The susceptibility of the patient to pneumocystis pneumonia could be attributed to the steroid therapy initiated for the original membranous nephropathy and the myelosuppression as a side effect of ganciclovir therapy.Our findings also show the relationship between CMV infection and pneumocystis pneumonia. Potentially, the pneumocystis pneumonia in our patient may have been related to the immunocompromised state induced by the steroid use and/or myelosuppression, which is an adverse reaction associated with ganciclovir therapy. Myelosuppression can be prevented by preemptive administration of ganciclovir, which can result in significant dose reduction compared to the normal dose .In conclusion, we report a case of oral ulcer caused by CMV infection in a patient with membranous nephropathy, which was severe and showed atypical manifestations. CMV infections in the oral cavity are not common. However, close collaboration with medical departments is essential when an immunocompromised patient with oral CMV infection is encountered, considering the possibility of a negative outcome.


The mineralogical composition of each sample was analyzed by X-ray diffraction (XRD) test, and the microstructure images of different samples were observed using scanning electron microscopy (SEM). The effects of the content of each component on the properties of the samples were discussed. Finally, the CO2 emission, energy consumption, and cost of producing fly-ash-based foamed geopolymer were analyzed. Overall, the material had the advantages of low energy consumption, low CO2 emission, environmental-protection ability, and waste utilization and thus has a broad application prospect in treating subsidence.The treatment of goaf subsidence is important for sustainable development. Geopolymer is a new type of cementing material with excellent mechanical properties, durability, corrosion resistance, and other advantages owing to its unique three-dimensional spatial aggregation structure. Herein, a type of preparation technology of fly-ash-based foamed geopolymer suitable for goaf filling was developed by adding a chemical foaming agent to the matrix of fly-ash-based geopolymer. The mechanical properties, chemical composition, and pore structure characteristics of the samples were discussed. When the samples with different contents of fly ash, sodium metasilicate, sodium stearate, H With the large-scale exploitation of mineral resources, the problem of goaf subsidence has increasingly become prominent ,2. If un2 emissions [2 emission materials [At present, researchers tend to use foamed concrete as filling material ,6,7,8,9.missions ,11,12. Tmissions ,14. Geopaterials ,19,20,21aterials ,26,27,28At the same time, with the development of the power industry, the fly ash emission of coal-fired power plants annually increases and has become one of the most widespread industrial wastes with large emissions in China ,34,35,362O2, and sodium stearate were prepared, and their physical properties, mechanical properties, mineral composition, microstructure, cost, and energy consumption were analyzed and discussed.Recently, fly ash has been used to synthesize geopolymer at room temperature after mechanical activation for 120 min . The relThe class F fly ash collected by the electrostatic sedimentation method in Fuxin thermal power plant  is used. According to the classification of fly ash in ASTM C618-17 , the CaO3. In this experiment, the foamed geopolymer samples with the target dry density of 420 kg/m3, cement content of 0–30%, and liquid–solid ratio of 0.43 were prepared. To study the influence of the content of different components on the properties of foamed geopolymers, samples with different mix proportions are designed according to On the basis of previous results , the dryThe cement, fly ash, sodium metasilicate, water, NaOH, and sodium stearate are weighed in accordance with the experimental design ratio. The entire preparation process is divided into four steps, namely, dry material mixing, activator configuration, foaming, pouring into the mold and curing .Dry material mixing, fly ash and cement should be pre-treated prior to use and placed into the oven for drying at 80 °C for 2 h to avoid moisture of raw materials. Fly ash and cement are poured into a cement mortar mixer and stirred for 5 min to mix evenly.Activator configuration, sodium metasilicate, NaOH, and sodium stearate are poured into 40 °C water, stirred for 2 min to fully dissolve, and then mixed evenly and set for 20 min.Foaming: the activator solution is poured into the cement mortar mixer and mixed with the dry materials, stirred for 3 min, mixed evenly, and set for 20 min, thereby allowing the activator solution to fully react with the fly ash in the dry materials.Pouring into the mold and curing: the hydrogen peroxide solution is poured into the mixer and rapidly stirred for 5 s. Then, the mixed slurry is poured into the 100 mm × 100 mm × 100 mm mold prepared in advance, set for 24 h prior to removing the mold, and finally placed into the constant temperature and humidity curing box for 28 d to obtain the required fly-ash-based foamed geopolymer cube sample. Then, the performance test is conducted.The phase composition of the sample was analyzed with a D8 advance X-ray diffractometer of Bruker Company. The X-ray diffractometer was D/max-γ B X-ray diffractometer, the radiation source was Cu Target Kα, the working voltage was 40 kV, current is 30 mA, and the wavelength was 1.5406 Å. The crystal structure and relative content were analyzed by scanning from 5° to 80° at 10°/min.The micro-morphology of the foamed geopolymer was observed using a FEI Quanta 250 FEG scanning electron microscope. The magnification was between 25 and 4000.WDW-300E electronic universal testing machine of Jinan Times Gold Testing Machine Co., Ltd.  was used to test the uniaxial compression performance of the samples to compare the uniaxial compressive strength of the samples with different mix proportions. The loading mode is strain controlled, and the loading rate is 0.2 mm/min. The compressive strength of the sample is obtained as follows:The average density of the three samples of the same mix proportion is obtained using the following calculation formula:The foamed geopolymer samples are rough, lusterless, and gray. The pore size of the sample surface is different, the distribution is irregular, and the pore size range is 1–8 mm. Uniaxial compression test was carried out to obtain the compressive strength and stress–strain curve of each sample.During the uniaxial compression experiment, the failure characteristics of the samples were observed a. The fiThe internal section of the sample after failure was observed. The findings indicate that most internal pore structures of the sample are closed, with different pore sizes, irregular distribution of pore structures, and have no orientation . Part ofThis analysis shows that the failure of fly-ash-based foamed geopolymer is primarily related to the “local large hole” in the sample and the bearing “weak zone” or “weak surface” formed by various initial defects.The uniaxial compressive strength of each sample is shown in The second type of curve is shown in The reaction process of geopolymers is a process of repolymerization after depolymerization. During this process, the fly ash spherical particles are dissolved, the Al-O and Si-O bonds are destroyed, and new non-crystalline gelatinous substances are formed ,52,53,542O3) and Si-O bond in fly ash under the action of alkali activator.The XRD patterns showed that the main components of the polymerization products were amorphous structures. It needs to be explained that obvious diffraction peaks appeared near 27 degrees in the spectrum, but this product was not the result of the polymerization reaction, but the quartz phase existing in the raw material fly ash, which is introduced by fly ash. There was a small amount of phosphate in the tap water for the test, and the aluminum phosphate produced by the ordinary Portland cement particles after absorbing the phosphate in the water. Metahewettite was formed by Cao in fly ash and cement under strong alkali environment.To further understand the pore structure characteristics and chemical reactions of the materials, the SEM images of the samples were compared and analyzed.The pore structure graphs and micrographs of group A samples are shown in The agglomeration of fly ash easily occurs after moisture absorption, which prevents the external alkali solution from entering and reduces the polymerization degree of fly ash. Few gelled polymers exist between spherical particles in Samples A1, A2, and A3, which cannot form an effective and firm connection, resulting in low density and compressive strength of foamed polymers.2O2 is different, the degree of polymerization of the obtained samples is also different. Relatively few gelatinous substances exist between fly ash particles of B1 samples, but the fly ash particles of B2 and B3 samples are almost filled with amorphous gelatinous substances. Correspondingly, according to the analysis in 2O2 content, the mixed slurry continuously has bubbles and move in the grout prior to the initial setting, thereby promoting the full flow contact of each component in the slurry and increasing the reaction degree of fly ash ball particles. On the contrary, when the H2O2 content is relatively small, the mixed slurry produces few bubbles prior to the initial setting, resulting in almost static slurry. The gelatinous substance produced by the first reaction is wrapped around the fly ash, preventing the ion exchange between the inside and outside of the coating. Moreover, the polymerization reaction cannot further enter the interior, resulting in a small quantity of polymerization material, which reduces the strength of the sample.2, which account for 7% of the world’s total CO2 emissions. Every 1 t of cement produced results in 1 t CO2 [2 emission from the production of 1 t of fly-ash-based foamed geopolymer is approximately 0.09 t given that no high-temperature treatment is required in the production process. Compared with ordinary Portland cement, producing the same quality of foamed geopolymer produces only 9% of the CO2 emissions of cement, thereby greatly reducing greenhouse gas emissions [Under the overall strategic layout of sustainable development implemented by the Chinese government, the greenhouse effect and global warming have prompted the people to actively develop low-carbon cement to replace ordinary Portland cement. In the 1990s, the world produced 1 billion tons of cement, resulting in 1 billion tons of CO 1 t CO2 . For flymissions ,57,58.Most industrial wastes contain aluminosilicate, such as kaolin, rock, fly ash, and other industrial wastes, which have been successfully used in the preparation of geopolymers. Therefore, the preparation materials of foamed geopolymers are widely available and low-cost. According to statistics, the ex-factory price of Portland cement is $1200–3200/t, and the production price of fly-ash-based foamed geopolymers is $200–700/t . Here isIn addition, as the production process of cement requires high temperature treatment process of “two grinding and one burning,” the energy consumption of cement production is extremely high, at approximately 3200–3500 MJ/t, but the energy consumption of fly-ash-based foamed geopolymer is only 800–1000 MJ/t.2 emission. Foamed geopolymer based on fly ash, H2O2, sodium stearate, sodium metasilicate, and NaOH was prepared. Uniaxial compression test results showed that the fly-ash-based foamed geopolymer prepared by this process presented brittle failure under uniaxial compression condition. Cracks appeared in the sample at the early loading stage, and cracks gradually broke through with increased load. The sample was also divided into several vertical bar shapes. The failure of foamed geopolymer was primarily related to the “local large pore” in the sample and the bearing “weak zone” or “weak surface” formed by various initial defects. The compressive strength of the samples with different component contents was significantly different. The test results showed that the compressive strength of the sample was between 0.221 and 2.221 MPa.A type of fly-ash-based foamed geopolymer, which is suitable for goaf filling is prepared. This material showed good bearing capacity, environmental friendliness, energy saving, and low COThe analysis of XRD experiment indicates that quartz was the residue of fly ash in the reaction product, Aluminum Phosphate is the crystal in the hydration products of cement hydration, and Calcium Silicate hydrates were the hydration products of Portland cement clinker during the cement hydration reaction.When the contents of each component differed, the reaction degree of the obtained geopolymer also differed. Relatively few gelatinous substances exist between the spherical particles of the fly ash of some samples, but the gaps around the fly ash particles of the other samples were almost filled with amorphous gelatinous substances, thereby resulting in high density of foamed geopolymer and corresponding high compressive strength.2 emission, energy consumption, and economy in the production of fly-ash-based foaming geopolymer were analyzed in detail. Compared with traditional foamed concrete, foamed geopolymer had the advantages of low carbon emission, low energy consumption, and low cost. In the future, it will have a wide application potential in goaf filling.Finally, the CO