Burnout is a common condition among health workers, characterized by emotional tiredness, depersonalization, and a sense of low personal accomplishment. Ethiopia has major health workforce management challenges, including shortages, poor motivation, retention, and performance, and research evidence is limited for health professionals' burnout status, particularly in Eastern Ethiopia. Therefore, this study is aimed at determining the prevalence of burnout and associated factors among health professionals working at governmental health facilities in Eastern Ethiopia.An institutional-based cross-sectional study was conducted among health professionals using structured self-administered validated questionnaires using the Maslach Burnout Inventory scale. Data were entered into Epi-Data version 3.1 and exported to SPSS version 22 for analysis. Multivariable logistic regression was used to determine the association between burnout and its predictors.Ethiopian Birr  and between 10,000 to 15,000 Ethiopian Birr , working in Pediatric Unit , and profession type  were factors significantly associated with burnout.A total of 508 health professionals were approached, out of which 501 participated (a response rate of 98.4%). The magnitude of burnout was 54.1 with a 95% confidence interval of 49.9–58.0%. Working in a hospital , age >/= 40  and 30–39 years , being female, being widowed , having intention of leaving work , using at least one substance , having a 6–11 years of experience , having no job supervision , monthly payment <10,000 Burnout affected more than half of the health professionals working in governmental health facilities in Dire Dawa. Health facility type, age, sex, marital status, intention to leave work, substance use, work experience, job supervision, monthly payment, profession type, and working unit were significantly associated predictors of burnout. Burnout is a condition that is thought to be caused by unmanaged long-term working stress . DepersoBurnout is becoming more common among healthcare providers, who have been classified as a high-risk category . EmployeBecause most studies on this topic are conducted in high-income nations, it is unclear how many people working in the health profession are burnt out globally . FurtherIn Sub-Saharan Africa, the health system suffers from a scarcity of healthcare professionals and overburdening of the available health professionals because of the increasing health-seeking behaviors in society, and physicians from Sub-Saharan Africa are migrating to the global workforce –11. Sub-In the last decade, Ethiopia has seen significant growth in primary health care services, including a large scale-up of health staff development and deployment. The number of health professionals in the population increased from 0.84 per 1,000 in 2010 to 1.5 per 1,000 in 2016. Ethiopia aims to meet the World Health Organization's (WHO) 2025 standard for Sub-Saharan Africa, which is a ratio of 2.3 health professionals per 1,000 people .Plenty of interventions have been implemented to lower the occurrence of burnout among healthcare professionals around the globe , 15. TheDespite these increments in burnout among health professionals, only a few studies have been conducted in Ethiopia. Most of these studies were conducted in hospitals, and on single health professionals , 17. FurA cross-sectional institutional study was undertaken in the Dire Dawa city administration's public health facilities from April 1 to April 30, 2020. Dire Dawa city is located in Ethiopia's eastern region, some 525 kilometers east of Addis Ababa, the country's capital. A total of 341,834 people live in the city administration. The city of Dire Dawa has 15 health centers, one general hospital, one referral hospital, and 768 health workers, according to the health office of the local administration .The source population included all health professionals who worked in the Dire Dawa City administration public health institutions. The study population consisted of all health professionals who were available at work during the study period and were included in the sample. This study comprised health practitioners who hadworked for more than 6 months in Dire Dawa City administration public health institutions prior to data collection. During the data collection period, however, health professionals who were extremely ill or unable to reply were omitted from the study.The study's minimum sample size was calculated by using both single population proportion formulas and a power approach based on results obtained from a similar study conducted in public hospitals in Amhara Regional State, Ethiopia . After tIn the city administration of Dire Dawa, there are two government hospitals and fifteen health centers. Each health facility's sample size was determined by proportionally allocating the calculated sample size depending on the number of health professionals on staff. The needed number of health professionals in each health facility was then picked using a simple random sample technique using the provided sampling frame  from each health institution's human resource department.Self-administered questionnaires adapted from previously conducted studies were used to collect data –24. The The measure used to assess burnout among health workers includes 22 questions organized into three burnout domains: 5 questions about depersonalization, 8 questions about personal accomplishment, and 9 questions regarding emotional exhaustion. Each question has a response ranging from 0 = (“never”) to 6 = (“daily”) based on a 7-point Likert scale. Scores in each of the three subscales of burnout were divided into high, average, and low categories based on the cut-offs used in previous studies . BurnoutEmotional exhaustion (EE) is defined as emotions of overextension as a consequence of one's job, with a score of 27 or more indicating high EE. Depersonalization (DP) is defined as emotional apathy and dehumanization of those who receive one's services, care, treatment, or instruction, with a score of 13 or more indicating high DP. Personal accomplishment (PA) is defined as sentiments of work stagnation, inefficiency, and underperformance, with a score of ≤ 31 indicating low PA –24.Bike primary hospital, which is one of the non-selected health institutions in Dire Dawa city, and the necessary changes were implemented accordingly. The data were examined for completeness, accuracy, and consistency by supervisors.The principal investigator conducted 2 days of training for data collectors and supervisors about the data collection technique and tool. A pre-test was conducted on 5% of health workers in the P-value of <0.25 in the bivariable logistic regression were included in the multivariable logistic regression analysis. A Hosmer-Lemeshow goodness-to-fit model was tested for model fitness. Finally, adjusted odds ratios with 95% confidence intervals were calculated to assess the strength of the association, and variables with a p-value of <0.05 were considered statistically significant factors.The data were compiled, cleaned, and coded before being entered into Epi Data version 3.1 and exported to SPSS version 22 for analysis. The mean, median, and percentage were used to summarize the data. Tables and figures were used to present descriptive data. Using a bivariable logistic regression, crude odds ratios with 95% confidence intervals were calculated to analyze the relationship between each independent variable and the outcome variable. Variables having a The Institutional Health Research Ethical Review Committee (IHRERC) at Haramaya University gave ethical approval. The city administration of Dire Dawa approved the conduct of this study and sent a formal letter of support to the respective health facilities in the City. Informed, voluntary, written and signed consent was taken from each study participant after explaining the aim and potential benefit of the study and confidentiality. This research was carried out in line with the Helsinki Declaration.Ethiopian Birr (ETB) monthly salary. Three hundred twenty-eight (65.5%) of the participants were urban residents  of the respondents were male, and the respondents' average age was 32 years old, with a standard deviation of 6.4 years. One hundred eighty-four (36.7%) of the participants were Muslim in religion, and 222 (44.3%) of the participants were married. In terms of educational level, 291 (58.1%) of professionals were bachelor's degree holders and 238 (47.5%) were nurses in the profession. Two hundred four (40.7%) of the participants had 6–11 years of experience, and 271 (46.6%) of the participants had ≤ 10,000 esidents .Four hundred forty-nine (89.6%) of participants had supervision in their current job, and 281 (56.1%) had ≤ 72 average working hours per week. Two hundred ninety-eight (59.5%) of the participants were not substance users. The majority of the participants, 290 (57.9%), worked at health centers, while 81 (16.2%) worked in the adult OPD. One hundred thirty-eight (27.5%) of the participants are currently working as—managers or have a position, and 166 (33.1%) had the intention of leaving their work .The prevalence of burnout was found to be 54.1% among health professionals working in Dire Dawa city administration public health facilities, with a 95% CI of 49.9–58.0%. Regarding components of burnout, 324 (64.7%) of the participants had high emotional exhaustion, 458 (91.4%) of the participants had high depersonalization, and 442 (88.2%) of the participants had low personal accomplishment .p-value <0.05 whereas age, education, residence, working area, and having a job position were not significant at p-value <0.05 but included in the multivariable analysis, based on reviewing previous studies , type of profession, intention of leaving a job, having job supervision, average working hours per week, facility type, sex, current marital status, residence area, substance use, total work experience, and current working unit had a significant association with health professional's burnout at  studies , 29, 32.In multivariable analysis, health professionals working in hospitals had a 3.55 times greater chance of burnout than those working in health centers . Participants over the age of 30 were more likely to have burnout, with >/= 40 years the most likely  than ages 30–39 . When compared to male health professionals, female health professionals had 2.41 times more chances of burnout . Widowed health professionals had a 3.39 times greater risk of burnout  compared to married health professionals.Burnout was twice as common among health professionals who planned to leave their employment  and those who used at least one substance  as it was among their counterparts. Health professionals who had 6–11 years of work experience had a 2.17 times greater risk of burnout compared to health professionals who had 5 years of experience or less . Health professionals who did not get job supervision had a 4.65 times higher risk of burnout than those who did .An average monthly payment of fewer than 10,000 ETB  and between 10,000 and 15,000 ETB  was a risk factor for burnout when compared to people who earn more than 15,000 ETB per month. In comparison to nurse professionals, midwives , medical laboratories , public health officers , and other health professionals  had higher burnout rates. Working in a pediatric unit increased the risk of burnout by 3.28 times compared to working in an adult OPD  .The magnitude of burnout among health professionals working in public health institutions in the Dire Dawa City administration was investigated in this study. Accordingly, the magnitude of burnout was 54.1%, which corresponds to the findings of a study in Iran (52.9%) , EthiopiIn this study, 64.7% of the respondents had high emotional exhaustion. This finding is consistent with the findings of research done at Debra Berhan University hospital (61.8%) . HoweverWhen comparing health professionals working in hospitals to those working in health centers, the risk of burnout was 3.55 times higher. This finding is congruent with a study conducted in Malawi . The reaThose in the age groups of 30–39 years and ≥40 years had 1.9 times and 3.98 times higher risk of burnout than those in the age group of below 30 years. This finding is backed up by findings from other studies done in the rural parts of Ethiopia . When coFemale health workers were 2.41 times more likely to experience burnout than male health professionals to experience it. This result contradicts the findings of research done at Gondar University hospital , and MalCompared to nursing professionals, midwives, medical laboratories, public health officials, and other health professionals, they had a higher risk of burnout. Compared to working in an adult OPD, working in a pediatrics unit had a higher risk of burnout. Such differences could be explained due to the nature of their profession and related workload. Those who had the intention of leaving work had around 2.28 times higher odds of burnout compared with their counterparts. This finding is in line with the study conducted in the Amhara region . This coThose who used at least one substance had 2.24 times the risk of burnout, and those who worked without supervision had 4.65 times the risk of burnout. This finding is supported by a study conducted in Malawi , 33–35. The odds of burnout were 5.69 and 2.74 times higher among health professionals who earn an average monthly payment of <10,000 ETB and between 10,000 and 15,000 ETB compared to those who earn above 15,000 ETB per month, respectively. A study conducted in Ethiopia backs up this finding . This coThere are certain limitations to this finding. Because it is a cross-sectional study, it can only reflect health professionals' experiences at the time of evaluation; hence, a causal relationship between health professional burnout and its predictors cannot be established. The other is that the study only included health professionals who worked in public health facilities. As a result, this may not be representative of health professionals that work in private health facilities.The findings revealed that the majority of health professionals working at Dire Dawa public health facilities had burnout. Working at the hospital, being 40 or more years of age, being female, being widowed, having an intention to leave work, using at least one substance, having 6–11 years of work experience, having no job supervision, earning an average monthly payment of <10,000 ETB and between 10,000 to 15,000 ETB, being a midwife, medical laboratory, and public health, and working in the pediatric unit were significantly associated predictors of burnout. We recommend the health facility manager focus on the female health professional's wellbeing to make them ready to give quality care. Establish mechanisms to improve staff retention; create different incentive mechanisms that increase staff income; and conduct regular supervision.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.The studies involving human participants were reviewed and approved by the Institutional Health Research Ethical Review Committee (IHRERC) at Haramaya University. The patients/participants provided their written informed consent to participate in this study.All authors contributed to the study's conception and design, as well as the acquisition, analysis, interpretation of data, drafting and revising of the article, gave final approval of the version to be published, agreed to submit it to the current journal, and agreed to be accountable for all aspects of the work.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Identifying selection on polygenic complex traits in crops and livestock is important for understanding evolution and helps prioritize important characteristics for breeding. Quantitative trait loci (QTL) that contribute to polygenic trait variation often exhibit small or infinitesimal effects. This hinders the ability to detect QTL-controlling polygenic traits because enormously high statistical power is needed for their detection. Recently, we circumvented this challenge by introducing a method to identify selection on complex traits by evaluating the relationship between genome-wide changes in allele frequency and estimates of effect size. The approach involves calculating a composite statistic across all markers that capture this relationship, followed by implementing a linkage disequilibrium-aware permutation test to evaluate if the observed pattern differs from that expected due to drift during evolution and population stratification. In this manuscript, we describe “Ghat,” an R package developed to implement this method to test for selection on polygenic traits. We demonstrate the package by applying it to test for polygenic selection on 15 published European wheat traits including yield, biomass, quality, morphological characteristics, and disease resistance traits. Moreover, we applied Ghat to different simulated populations with different breeding histories and genetic architectures. The results highlight the power of Ghat to identify selection on complex traits. The Ghat package is accessible on CRAN, the Comprehensive R Archival Network, and on GitHub. Many important traits in plants, animals, and humans are polygenic and depend on the cumulative effects of many loci, each contributing a small proportion of the total genetic variation . StudyinRecently, large sample sizes, especially as utilized in human genetic studies, have allowed a more powerful understanding of the genetic architecture of polygenic traits using genome-wide association studies (GWAS) . After cQPC, which detects adaptive divergence while controlling for population substructure. Conditional QPC detects adaptive divergence based on the excess of variation observed among populations versus variation observed within populations  ” and canParallel-Package {parallel}: Support for parallel computation, to speed the computation time in large data analysis (rrBLUP-Package {rrBLUP}: Ridge regression for estimating marker effects (RR-BLUP) .These may be installed and loaded before running Ghat by typing: >install.packages) and then typing > library and > library(rrBLUP) , step-1.For its simplest implementation, Ghat requires only 2 vectors of input. The first of these is a vector of allele substitution effects, and the second is a vector containing the allele frequency change between the 2 generations   are required , step-4 To demonstrate the power and flexibility of the Ghat R package, we used it to test for selection during the past 50 years of wheat breeding in Western Europe. To achieve this, we leveraged a dataset representing 50 years of commercial winter wheat varieties that were previously published by Voss-Fels and colleagues  biomass parameter traits, including above-ground biomass, kernels per sqm, and thousand-kernel weight; (3) quality parameter traits, including crude protein, protein yield, and nitrogen use efficiency; (4) morphological traits, including plant height, green canopy duration, and kernel per spike; and (5) disease resistance traits, including resistance to powdery mildew and resistance to stripe rust. All cultivars were genotyped with a 15K SNP Illumina Infinium iSelect genotyping array . All traits were tested in 3 different environments by Pearson correlations between phenotype and the year of release  , we can spearman correlations between the Ghat results from 3 and 50 years of phenotypes were estimated. A significant  correlation was found between Ghat values from 3 and 50 years of phenotypes, indicating that even in the absence of historical phenotypic information, Ghat can be used to understand historical patterns of selection.To dissect the history of selection in wheat more precisely, we divided the 50 years of breeding into 10 5-year time periods and applied the Ghat test successively for each period. We excluded the green canopy duration trait from this analysis since it was only measured under 1 environmental condition. For the remaining 14 traits, we calculated the average phenotypic value across the 3 different environmental conditions. For example, during the period from 1966 to 1975, significantly selected traits according to the Ghat test included grain yield, harvest index , above-g2h), sample size (n), trait polygenicity [number of QTL (nQTL)], and marker density (MD) on the performance of Ghat.To complement the above wheat analysis, we additionally generated a simulated cattle dataset with divergent selection to further test and demonstrate the efficacy of Ghat. This allowed us to evaluate how the Ghat package performs in terms of different genetic architectures and experimental parameters, which are never perfectly known using real data. In this analysis, we assessed the impact of trait heritability  in each dataset. Then we measured the P values coming from using these different datasets with different n. Group 2 was simulated to investigate the effect of h2 on Ghat performance. For this, we simulated 10 datasets with a constant n, nQTL, and MD, but with a different h2 (from 0.1 to 0.95) in each dataset. Group 3 investigated the effect of nQTL on Ghat performance. For group 3, we simulated 10 datasets with a constant n, h2, and MD, but with a different level of nQTL  in each dataset. Group 4 was simulated to investigate the effect of MD on Ghat performance. In this group, we simulated 10 datasets with a constant n, h2, and nQTL but with a different level of MD (from 30 to ∼ 1 million markers) in each dataset.Four groups of datasets were simulated for studying the performance of Ghat . Each grhttps://github.com/Medhat86/Selection-and-adaptation-on-simulated-bovine-genome).Genomic prediction methods like BayesR, BayesB, and BayesC assume that the allele substitution effects follow a distribution in which many effects are zero and some have moderate effects, subsequently these methods may for some genetic architectures lead to higher accuracy than the GBLUP/rrBLUP methods . The effn, h2, nQTL, and MD) to evaluate the detection accuracy by evaluating true discovery rates and false positives. Confidence intervals were obtained from P-values of the same traits from the Ghat permutation test using the formula from Ghat is calculated as the summation of the estimated effect of every SNP scored multiplied by its effect size, according to model 1. We implemented identical analyses on selected and unselected populations across 1,000 simulation replications for 4 different tests  under selection. Conversely, in the selected population 99.3% of replicated traits found highly significant under selection . The P-value also shows a decreasing trend with increasing n  in the selected population. Moreover, the confidence interval decreases with increasing n.Based on 1,000 replications, h2 on the power of Ghat. Higher h2 led to a more powerful test. When we tested the unselected population with a constant n = 400, nQTL = 150 and 10,020 markers, 100% of replicates were not significantly  under selection. In the selected populations 100% of replicates were significant for selection . The P-value shows a consistency decrease with increasing trait h2 (from 0.1 to 0.95 of h2).Based on 1,000 replications, nQTL on the power of Ghat. When we tested the unselected population with a constant n = 400, h2 = 0.5 and 10,020 markers, 99.7% of replicated traits were not significantly  under selection. In the selected population, 100% of replicated traits found highly significant selection . The power of the test increased with increasing the nQTL in the selected population. This can be restated to emphasize that, all else being equal, Ghat is more able to detect selection on traits that are more complex (i.e. controlled by a large number of QTL).Based on 1,000 replicates, P-values of the Ghat test for selection. When we tested the unselected population with a constant n = 400, h2 = 0.5 and 150 QTL, 90% of replicated traits were not significantly  under selection. While, in the selected population 100% of replicated traits were highly significant for selection . The P-value also shows an inconsistent increase of significance along with an increasing MD in the selected population (from 30 to ∼ 1 million MD). This may be caused by multicollinearity in the data when there are a high number of markers  and relatively small n .Based on 1,000 replications, https://cran.r-project.org/package=Ghat) for the 3 major operating systems: MS Windows, Linux/Unix, and Mac OS. We have also released the source code on GitHub (https://github.com/Medhat86/Ghat) so that users can compile for specific platforms or modify the code as needed.We have released Ghat package on CRAN  relative to current high-density marker panels and/or whole-genome sequencing approaches, so this analysis did not represent all the challenges associated with implementing Ghat in these settings. In particular, high-density panels include markers that are in strong LD, which must be properly accounted for in any Ghat implementation.To address this, we used a simulated SNP panel to further explore the utility of Ghat when high LD between markers is present. We found that Bayesian methods, such as BayesC , 2011. BUnlike methods for detecting selected loci, Ghat can quantify the direction of selection even in the absence of phenotypes from a preselection population. For breeders and breeding organizations, this represents a practical solution for tracing selection and adaptation history based on phenotypes measured from modern germplasm coupled with DNA samples saved from the past. From an economical perspective, this may allow a retrospective evaluation of traits that were inadvertently selected in the past, and therefore may productively serve as the targets of intentional selection in the future. Advances in phenotyping technologies make measuring novel traits increasingly possible (The Ghat test and implementation via the package described herein does not require immense sample size, individual genotypes, and/or genome-wide significant SNPs, as demonstrated by our implementation with a relatively modest sample size of 191 wheat individuals genotyped for 8,710 SNPs. Notably, this approach is more powerful when traits are controlled by a large number of QTLs (jkac319_Supplementary_DataClick here for additional data file.


Moyamoya syndrome (MMS) is a cerebrovascular disease characterized by stenosis of the internal carotid arteries and the formation of an abnormal vascular network at the base of the brain. MMS usually occurs secondary to various conditions, particularly Down syndrome, and sickle cell anemia, and presents with motor deficits, sensory symptoms, recurrent ischemic strokes, hemodynamic transient ischemic attacks, recurrent seizures, and hemorrhage. Trisomy 13 (Patau Syndrome) is a chromosomal abnormality that may be characterized by full or partial trisomy of chromosome 13. Phenotypic features of partial trisomy 13 include leukoencephalopathy, hippocampal hypoplasia, intellectual disability, facial anomalies, and others. Herein, we report a case of a 19-year-old female diagnosed with partial trisomy 13q, characterized by two large duplications in the 13q14 and 13q31 regions, with trisomy-induced bilateral MMS – the first known case to be discussed in literature. Particularly, our patient was identified to have a gain of 22Mb within the 13q14.11q21.31 region – a duplication that has not been described previously. Our patient suffered four strokes between the ages of 5 and 7, later developing intractable seizures, hemiplegia, spasticity in all limbs, global delay, and regression. Despite bilateral encephaloduroarteriosynangiosis and being on several antiepileptic medications, the MMS continued to progress, confounded by the partial trisomy 13. Studies must elucidate the association between mitochondrial damage and MMS, as well as mechanisms of epilepsy associated with chromosomal abnormalities, particularly in the context of underlying mitochondrial diseases. MMS is Trisomy 13 (Patau Syndrome) is an uncommon and severe chromosomal abnormality caused by a trisomy of chromosome 13 and is associated with a prevalence rate of 1.68/10,000 ,8. PossiThe mechanisms leading to the characteristic symptoms of trisomy 13 are not yet well defined. However, generation of reactive oxidative species (ROS) seems to play a key role in its pathogenicity . StudiesMitochondrial abnormalities are also implicated in neuronal damage, as neurons are mostly dependent on mitochondria in oxidative stress conditions . These mHerein, we report a case of a 19-year-old female diagnosed with partial trisomy 13q, characterized by two large duplications in the 13q14 and 13q31 regions, with trisomy-induced mitochondrial dysfunction, presenting with bilateral MMS. To the authors’ knowledge, this is the first article reporting an association of trisomy 13 with MMS. This case report is reported in accordance with the CARE guidelines .2Our patient, a young Saudi female, is the youngest child of non-consanguineous parents, and has two older siblings who are healthy. She first presented to our hospital at the age of 6 years with a history of bilateral frontal stroke and left partial stroke. Her first stroke occurred when she was less than 6 years old, presenting with right-sided weakness and seizures. The seizures began as absence seizures and eventually progressed to tonic seizures. The patient was managed with antiepileptic drugs and blood thinners. Radiology did not reveal arteriovenous malformation or aneurysm. The patient had her second stroke a few months later, which resulted in more severe right-sided weakness. Moyamoya syndrome was suspected due to the recurrent cerebrovascular accidents, and a magnetic resonance angiography (MRA) a and b aLeft-sided encephaloduroarteriosynangiosis (EDAS) was performed three months later, but the patient's seizure activity persisted. Thus, she was diagnosed with epilepsy, and was started on prescription antiepileptic drugs, namely valproic acid, clobazam, and levetiracetam.The patient was then brought to our hospital, where she was scheduled for right-sided EDAS. However, she suffered a third stroke involving the right middle cerebral artery while awaiting the procedure a–c. MRA Despite undergoing EDAS on both sides, the patient's condition did not improve, and she continued experiencing intractable seizures. She was continued on valproic acid, levetiracetam, and clobazam. The patient developed bilateral facial palsy and was no longer able to tolerate oral intake without risk of aspiration. At the age of 7, Nissen fundoplication and gastrostomy were performed five months after right-sided EDAS to allow easier feeding.By the time the patient was 14 years old, she had developed global developmental delay and regression, uncontrollable seizures, and was on multiple antiepileptic medications. Brain MRI performed at age 14 showed scattered old ischemic insults with laminar necrosis, gross parenchymal loss, and gliotic changes with ex vacuo dilatation of the ventricles. No acute intracranial hemorrhage or acute infarction was seen. Further investigations revealed uncorrectable hyponatremia , constipation, precocious puberty, and scoliosis. Correction of hyponatremia to the level of no seizure activity became the aim of treatment, however, did not prove to be successful.Additionally. high lactate levels were detected in the venous blood gases of the patient. Thus, magnetic resonance spectroscopy was performed at an external hospital, and small lactate peaks were identified in the basal ganglia regions. Unfortunately, this image could not be retrieved currently for the purpose of this case report. However, given the patient's history of recurrent strokes, this led us to suspect mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS) syndrome. Comparative genomic hybridization (CGH) was performed to rule out this condition and revealed partial Trisomy 13.Whole exome sequencing (WES) performed when the patient was 17 years old revealed several mitochondrial diseases among others, seen in 1.A gain of 22Mb within the 13q14.11q21.31 chromosomal region, determined to be likely pathogenic, non-distal trisomy 13q: The proximal long arm of chromosome 13 is partially duplicated, resulting in a highly variable phenotype. While the exact nature of this duplication has never been described in literature, similar duplications have been related to partial trisomy 13q in the past. Growth and developmental delays accompany this phenotype, as well as craniofacial dysmorphisms .2.A gain of 24Mb within the 13q31.3q34 chromosomal region, determined to be pathogenic, distal trisomy 13q: In distal trisomy 13q, the distal long arm of chromosome 13 is partially duplicated, again resulting in a variable phenotype. In addition to microcephaly and holoprosencephaly (incomplete development of the forebrain), this phenotype may be characterized by intellectual disability, psychomotor delay, and craniofacial dysmorphism .Chromosomal microarray analysis (CMA) performed when the patient was 19 years old identified two large duplications that were consistent with a genetic diagnosis of partial 13q trisomy  1.A gainMendeley Data, V1, doi: 10.17632/7b8nf7gp3x.1 [All WES and CMA data from this patient can be found on a dataset on f7gp3x.1 . A full 3MMS is a cerebrovascular condition characterized by stenosis of the ICAs causing recurrent strokes, TIAs, and seizures . It has MMS is a widely recognized cause of stroke in children and must be considered among the differential diagnoses of any child presenting with symptoms of cerebral ischemia. It can be identified on brain CT or MRI by the visualization of small collateral vessels growing in the bottom of the cerebrum, resembling a “puff of smoke”. A CT scan may also reveal hemorrhage and atrophy of the affected hemisphere . Early oTrisomy 13, or Patau Syndrome, is a chromosomal aberration that results in an extra duplicate of chromosome 13 being present in the body cells. Trisomy 13 may be full trisomy 13 , mosaic trisomy 13 (a third copy of chromosome 13 exists only in some cells), or partial trisomy 13  ,23. PartWe report a case of bilateral MMS occurring in a 19-year-old patient, secondary to underlying partial trisomy 13q. To our knowledge, this is the first recorded case of the two syndromes coexisting in a patient in literature. We believe her partial trisomy 13q has undue influence on her prognosis and improvement, due to the associated oxidative stress and mitochondrial illnesses. This matter has never been discussed previously, making it imperative that further research is conducted to identify the possible consequences of trisomy 13 on MMS.While little to no data exists on the association between trisomy 13 and MMS, it has been established that the pathogenic changes seen in trisomy 13 are a direct cause of oxidative stress, with the faulty genes being primarily involved in the redox balance regulation . The priFurthermore, Alramadan et al. established that 73% of pediatric moyamoya patients who underwent EDAS had improved seizure outcomes after surgery. Six out of eleven patients also became seizure-free . Furtado4In conclusion, 13q14 and 13q31 are crucial chromosomal regions that may be associated with oxidative stress and mitochondrial abnormalities leading to the development of MMS. We present a case of partial trisomy 13q that led to mitochondrial disease and ECFC dysfunction, leading to the development of MMS. Unfortunately, despite bilateral EDAS, the patient's MMS continued to progress, resulting in refractory seizures and spastic quadriplegia. MMS should be suspected and investigated in all children presenting with stroke. Greater research is needed to delineate the clinical features of MMS refractory to EDAS. Partial trisomy 13 is a significant chromosomal abnormality and studies should aim to shed more light on its varying presentation and genetic expression and elucidate the mechanisms of epilepsy associated with chromosomal abnormalities, particularly in the context of underlying mitochondrial illnesses. The number of studies demonstrating how mitochondrial damage is implicated in trisomy disorders is limited and greater research must be conducted to establish preventative measures, such as early screening tests for moyamoya disease. Lastly, we urge clinicians to suspect underlying cytogenetic causes in all pediatric patients presenting with recurrent strokes and seizures, hasten diagnosis, and begin appropriate treatment before further disease progression.Written informed consent was obtained from the patient's parents for publication of this case report and accompanying images. Patient anonymity is maintained throughout this manuscript.All authors listed have significantly contributed to the investigation, development and writing of this article.This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.https://doi.org/10.17632/7b8nf7gp3x.1.Data associated with this study has been deposited at Mendeley Data, V1, The authors declare no competing interests.


Microalgae are a kind of photoautotrophic microorganism, which are small, fast in their growth rate, and widely distributed in seawater and freshwater. They have strong adaptability to diverse environmental conditions and contain various nutrients. Many scholars have suggested that microalgae can be considered as a new food source, which should be developed extensively. More importantly, in addition to containing nutrients, microalgae are able to produce a great number of active compounds such as long-chain unsaturated fatty acids, pigments, alkaloids, astaxanthin, fucoidan, etc. Many of these compounds have been proven to possess very important physiological functions such as anti-oxidation, anti-inflammation, anti-tumor functions, regulation of the metabolism, etc. This article aimed to review the physiological functions and benefits of the main microalgae-derived bioactive molecules with their physiological effects. Their single frond cannot be divided into rhizomes and leaves [Microalgae are a class of autotrophic single-celled organisms that are widely distributed on land and in oceans, and they are rich in nutrients and active in photosynthesis. Moreover, they are also one of the oldest organisms on earth . At the d leaves . Accordid leaves . Microald leaves . Anotherd leaves .Microalgae contain a large number of nutrients. The protein content in microalgae can reach 2.4 times that which is in cheese, three times that which is in pork, four times that which is in flour and fish, and five times that which is in eggs . ProteinHaematococcus pluvialis is rich in astaxanthin [Seaweed and Microchlorococcus. Here, we summarize some of the activities and benefits of microalgae-derived extracts and biomolecules.Some microalgae contain a large amount of polysaccharides, which usually have the functions of anti-tumor, anti-virus, anti-aging, enhancing immune activity, anti-radiation, anti-mutation, and they regulate blood lipid levels . Other maxanthin . MoreoveMucidosphaerium sp. Inhibits the inflammatory responses in the skin and joint cells, which attributed to its antioxidant properties. In dermal fibroblasts, dermal papilla cells, or synoviocytes, treatment with the extract can down-regulate the expression levels of interleukin-1β (IL-1β), IL-6, and tumor necrosis factor-α (TNF-α), which are cytokines that promote inflammation. Moreover, the extract was able to suppress the mRNA expressions of cyclooxygenase 2, matrix metalloproteinase 1 (MMP-1), MMP-3, and MMP-9, which are enzymes that are involved in extracellular matrix remodeling. This is associated with the reduction of total MMP protease activities. The intracellular reactive oxygen species (ROS) levels in synoviocytes that were treated with the extract from Mucidosphaerium sp. was reduced, and the antioxidant activity was estimated as 178.3 ± 0.9 μmol trolox equiv/g, which was higher than that in the control cells [Microalgae produce many molecules with antioxidant activities. The extract of ol cells .diatoms, dinoflagellates, and golden algae, contain a high content of fucoxanthin, an orange-colored pigment, which has a strong antioxidant effect. It can eliminate free radicals and reduce the level of cellular oxidative stress, producing significant antioxidant effects [Many microalgae, such as  effects . As a re effects ,16.Chlamydomona, a genus of Antarctic ice microalgae is a natural marine resource that is rich in polysaccharides. In mice that are treated with polysaccharides which are derived from Chlamydomona, the oxidative stress in the internal organs such as the liver and kidney and the serum markers of tissue damages such as amino transferases were significantly reduced. The serum levels of glutathione and glutathione peroxidase, and the contents of nitric oxide (NO) and malondialdehyde in the spleen and liver were also decreased after the polysaccharide treatment. All of these changes are associated with increases in the expression level of genes that are involved in the clearance of ROS, such as manganese superoxide dismutase, catalase, etc., and decreases in those involved in NO production, such as inducible NO synthase in the tissues and cells of those treated animals [ animals .Auricularia terrestris. At 10 mg/mL, this polysaccharide extract demonstrated 92.7% of the scavenging activity against hydroxyl radicals, and a 0.445 reducing power for potassium ferricyanide [Nostoc Sphaeroides has been shown to perform a scavenging activity that is used to eliminate hydroxyl and super anion free radicals and hydrogen peroxide in a dose dependent manner. In, in vivo experiments, the phycocyanin treatment can significantly reduce the production of malondialdehyde and the content of peroxides in the blood and liver and protect cell membranes and red blood cells [B. braunii, a well-known microalga, can secrete a large amount of exopolysaccharide (EPS), which is a heteropolysaccharide with a high molecular mass, consisting of about 7–9% uronic acid, 2–4% protein, and 1.5–2% sulfate moieties. The monosaccharide composition of EPS is 52–54% galactosyl, 35–36% glucosyl, 9–10% arabinosyl (9.41–10.32%), and 1.8–2% fucosyl moieties. The crude EPS extract without deproteinization performed a stronger degree of antioxidant activity than the purified polysaccharide did. The 50% inhibitory concentration (IC50) of the crude EPS extract, which is the concentration at which half of the hydroxyl radicals are inhibited, is about 1.67 mg/mL, which is significantly lower than that of the deproteinized EPS, which has an IC50 of about 3.04 mg/mL [Wang et al. in 2014) extracted 96.7% water-soluble polysaccharides from icyanide . Phycocyod cells . B. brau14 extracCalothrix strongly inhibited the production of phenyl-1-picrylhydrazine free radicals. The IC50 of this ethanol extract is about 30 μg/mL. Babić et al. (in 2016) found that the ethanol extract of Phormidium can perform about 23 g/kg antioxidant activity, which is the highest in the plasma iron reduction ability test. It contains 12 kinds of polyphenols, mainly phenolic acids and flavonoid glycosides. The contents of the quinic acid, gallic acid, and vanillic acid are 502, 84.9, and 50 μg/g, respectively, which may be the main reason for its strong antioxidant activity. Botryococcus braunii releases significant quantities of EPS, which can be used in the preparations of regular and medicinal foods, and cosmetics. To obtain EPS, B. braunii SCS-1905 was extracted and precipitated in ethanol, and then, it was deproteinated. After that, column chromatography using a DEAE-cellulose matrix was used to purify the EPS [The extract from  the EPS .Haematococcus pluvialis is a species of microalgae that is identified in fresh water. It is in the family Haematococcaceae, of the order Chlamydomonadales. This unicellular green alga can synthesize significant amount of carotenoids, which can achieve to 2–5% of the dry mass. The astaxanthin content in haematococcus pluvialis is very high. For this reason, Haematococcus pluvialis has the drawn attention of scientists who are interested in extracting and purifying naturally derived astaxanthin from this organism [organism . As a caorganism . The antorganism .The antioxidant effect of microalgae is one of its aspects that was identified the earliest and studied the most extensively. The antioxidant mechanisms include the direct scavenging of free radicals, the chelation of pro-oxidative metal ions, the regulation of ROS-producing oxidase, and the enhancement of oxidative defense system. This well-known antioxidant function of the microalgae has the most mature application in the market. For example, astaxanthin is currently one of the most popular products that is derived from microalgae. Since oxidation occurs in every part of the body, it is related to the development of many diseases. However, information showing the antioxidant activities of microalgae products, improvement of disease progression, and the underlying molecular mechanisms is still limited. More in-depth research on the antioxidant effects of microalgae is anticipated in the future.Spirulina microalgae on glioblastoma have been studied in vivo. The brain samples of rats that were treated with Sprirulina microalgae were analyzed for the tumor size and gene expression levels. It appeared that the Spirulina treatment inhibited the tumor cell proliferation and induced cell death. In addition, the spirulina treatment significantly up-regulated the levels of miR-34a and miR25b micro-RNA that play a role in the PI3K/Akt/mTOR signaling transduction pathway [Nodularia and Anabaena. NoA induced the apoptosis of cancer cells in a caspase-dependent manner. NoA at 0.7–4.5 μmol/L triggered cell death in several human cancer cells such as lung cancer A549 cells and H460 cells, pancreatic cancer BxPC cells, breast cancer MCF-7, JIMT1 and SKBR3 cells, glioma U87 and U251 cells, and ovarian cancer OVCAR5 cells. This antiproliferative activity of NoA was more potent in p53 mutant cell lines, suggesting the presence of a novel mechanism that is mediated by NoA, which could be a drug target for the treatment of tumors, independent of the p53 pathway [The direct anti-tumor effects of  pathway .Candida oryzae were cytotoxic to breast cancer MCF7 cells and colon cancer HCT116 cells, which had IC50 numbers at 47.8 to 232.0 mg/L [Anabaena sp. can induce apoptosis in cells of acute myeloid leukemia [Euglena gracilis extract significantly attenuated the lung tumor growth, possibly by altering the gut microbiota. It significantly increased gut microbiota diversification, decreased the Firmicutes to Bacteroidetes proportion, and promoted the abundance of Akkermansia and Muribaculum [Pham et al., found that extracts from three strains of 2.0 mg/L . Additioleukemia . Studiesibaculum .C. raciborskii CYP011K and Nostoc sp. perform an inhibitory activity in CT-26 and 3LL cancer cells [Chlorella EPS was shown to inhibit the growth and the reduce viability of Hela cells, a classic cervical cancer cell line. The EPS treatment regulated the expression levels of differentially expressed genes and activated various cancer and apoptosis-related proteins, which include mitogen-activated protein kinase (MAPK), TNF-1α, PI3K, and Akt. The changes in the expression levels of 13 proteins that are involved in signal transduction pathways were found in cells that were treated with Chlorella EPS [Silva-Stenico et al., screened the anticancer activity of extracts that were obtained from 24 strains of microalgae, using CT-26 and 3LL cancer cells. The methanol extracts (MEs) of er cells . Chlorelella EPS .Dunaliella sp. These microalgae were allowed to grow in normal or stressed conditions, and then were extracted using different methods. The anti-tumor activities of the extracts were evaluated in 4T1 murine breast cancer cells. The experimental results demonstrated that the water extract performed a higher level of cytotoxic activity than the ethanol and hydroalcoholic extracts did. The extract from Dunaliella sp. which was cultured in a stress condition performed a higher level of anti-tumor activity than it did in a normal condition. This cytotoxic mechanism is apoptosis, not necrosis, due to the presence of DNA fragmentation and caspase-3 activation. The direct injection of the extract from the stressed Dunaliella sp. inhibited the tumor growth and enhanced the immune function in zebrafish. All these results were associated with the decreases in the expression levels of Arg-1, NOS-2, and COX-2 genes, which play roles in immunomodulation [Lazado et al. (in 2019) investigated the anticancer activity of the extracts from microalga dulation .Due to their diversity of their distribution, habitat conditions, and species, the probability of identifying anti-tumor substances from microalgae is relatively high. In fact, many active compounds have been isolated from them. In the meantime, with the development of bioreactors, cell cultures, and bioengineering and other technologies, the research on the anti-tumor effects of microalgae has made meaningful progress, recently. However, it is still in the initial stage of screening for anti-tumor substances and their functional mechanisms. Additional well-designed clinical trials should be conducted to evaluate the efficacy and safety of bioactive compounds from microalgae such as polysaccharides in patients with cancers.Chlorella, and analyzed the characterized activities of the fraction 4 (CEP4). CEP4 is made of sulfated heteropolysaccharides, which is composed of 41% glucosamine hydrocholoride, and 21% glucuronic acid moieties. In TAW264.7 macrophage cells, the CEP4 treatment significantly increased the production of cytokines such as TNF-α and IL-6, and NO. The effects were found to be associated with the increase in the cytokine gene expressions. In addition, the CEP4 treatment reduced the expression levels of receptor-interacting protein kinase 1 (RIPK1) and toll-like receptor 4 (TLR4), suggesting the reduction of TLR-signaling transduction. Other pathways mediating cytoplasmic DNA sensing, and lectin receptor (C-type) signal transductions were also affected. A further investigation of protein–protein interactions identified the key player proteins mediating the immunomodulation roles of CEP4 [Tribonema sp. by Chen. The structure and function of this sulfated polysaccharide were studied. This heteropolysaccharide includes a significant proportion of the galactosyl group and exhibits immunomodulatory activities. Its treatment activates macrophages and stimulates the production of cytokines such as IL-6, IL-10, and TNF-α [Wu et al. (in 2021) isolated, fractionated, and purified the extracellular polysaccharide (CEP) of  of CEP4 . A sulfand TNF-α .Scenedesmus acutus in the presence of higher concentrations of K2HPO4 and MgSO4, and obtained an EPS with a higher glucose content than those that were cultured at lower K2HPO4 and MgSO4 concentrations. This EPS had immunostimulatory properties and stimulated the proliferation of human peripheral blood mononuclear cells (PBMCs) [Chlorella ellipsoidea and subsequently separated the polysaccharides using anion exchange chromatography to obtain fractions for further analysis of their molecular characteristics and immunomodulatory activities. The results showed that some factions of the polysaccharides could stimulate productions of NO and cytokines in RAW264.7 macrophage cells. These fractions activated MAPK and nuclear factor-κB pathways, and in turn, stimulated the expressions of the cytokine mRNA [Patwal (in 2021) cultured  (PBMCs) . Qi et aine mRNA .Dunaliella salina secrete EPSs. The extraction, which was obtained using ethyl acetate, resulted in a fraction that modulated the proliferation rates of human PBMCs and RAW 264.7 macrophages in a healthy human being. The treatment with this ethyl acetate fraction enhanced the production of cytokines such as interferon-γ (IFN-γ) and TNF-α. The effects on cell proliferation are bimodal. At 250 and 500 mg/L, the fraction inhibits the cell proliferation, whereas at 1000 and 1500 g/L, it promotes the cell proliferation. When the PBMCs were treated with the fraction at 750 and 1000 mg/L, the TNF-α production was increased, suggesting that it has an immunostimulatory function. In RAW 264.7 macrophage cells, this faction inhibited cell proliferation with an IC50 of 691 mg/mL, and NO production with an IC50 of 630 mg/mL [30 mg/mL .Schizochytrium in dairy goats, which had a regular diet of alfalfa hay and concentrate. The goats were separated into four groups, a control group with the concentrate without the addition of Schizochytrium, and three experimental groups that were fed 20, 40, or 60 g of Schizochytrium sp. in concentrate, per day. The blood samples were collected from the animals that were fed Schizochytrium for 20, 40, and 60 days and they subjected to the analysis of monocytes and neutrophils. Compared with those in the control group, the mRNA levels of the components in cytokine signal transduction pathways such as MYD88 and TLR4, pro-inflammatory cytokines such as IL-1β and TNF-α, and chemokines such as CCL5 and CXCL16 in monocytes of the supplementary groups were all reduced. Interestingly, when it was compared with those of the controls, the mRNA levels of MAPK and IL-1β in the neutrophils of the animals that were fed 40 and 60 g/d Schizochytrium were increased, whereas that in the group that was fed 20 g/d, it was decreased. These results indicate that Schizochytrium can modulate immunity in goats [Alexandros (in 2021) investigated the effects of dietary supplementation with in goats .Chlorella powder on the cyclophosphamide-induced immunosuppression have been studied in mice. The results showed that Chlorella powder significantly increased the proliferation of lymphocytes, the phagocytosis of macrophages, and the cytotoxicity of natural killer cells. In addition, the treatment decreased the plasma levels of cytokines such as IL-12 and IFN-γ, which are associated with histological improvements in the spleen of cyclophosphamide-treated mice [Chlorella powder in humans, and they have reported that the oral administration of Chlorella powder for 4, 5, and 6 weeks significantly increased the secretory immunoglobulin A levels of the saliva [Micractinium simplicissimum, an Antarctic freshwater microalgae, and studied the effects of this extract on inflammation in RAW 264.7 macrophages cells. The treatment with the ethanol extract reduced the expression levels of IL-6, cyclooxygenase (COX)-2, inducible NO synthase, and TNF-α, suggesting that it can perform anti-inflammation activities. This was also associated with reduced NO production [The effects of ted mice . Others e saliva . Chae usoduction .Tisochrysis lutea F & M-M36 and compared the effects of this ME with that of the equivalent concentration of fucoxanthin in RAW 264.7 macrophage cells that were treated with lipopolysaccharide (LPS). The treatment with this ME significantly inhibited the COX-2-dependent prostaglandin E2 (PGE2) production when it was compared with the control group. Both the ME and fucoxanthin showed the ability to stimulate the expression levels of IL-10 and heme oxygenase-1 (HO-1) and inhibit IL-6 and arginase 1 in those cells. Additionally, the ME treatment led to an increase in the expressions of NLR family pyrin domain-containing 3 (NLRP3) and mir-223, and a decrease in mir-146b when it was compared with the fucoxanthin treatment. These results indicate that the ME of Tisochrysis lutea F & M-M36 has anti-inflammation functions, which can be attributed to the regulation of PGE2 production, and the expression levels of genes that are involved in immunity such as NLRP3 and mir-223. The active components in the ME, probably, are compounds with phenolic structures, which may work together with fucoxanthin to regulate inflammation [Bigagli used methanol to extract ammation .Microalgae polysaccharides play important roles in immune regulation. Previously, studies of polysaccharides have been mainly focused on terrestrial organisms and macroalgae such as brown algae and red algae. Studies of polysaccharides from microalgae have been limited. Microalgal polysaccharides are abundant and structurally diverse, which leads to them performing distinct physiological activities. More studies of the relationships of the structures and these activities will be of great value.chlorella, which showed antibacterial and anti-autotoxic functions [Staphylococcus epidermidis, Staphylococcus aureus, and Streptococcus agalactiae, but it did not perform a bacteriostatic activity against strict anaerobic bacteria [mycobacterium tuberculosis by inhibiting the UDP-galactopyranose mutase and arylamine-N-acetyltransferase activities, demonstrating the great potential of fucoxanthin for the treatment of tuberculosis [Polysiphonia denudota and P. denudataf fragilis, and found that they performed antibacterial activities, strongly [Molecules that perform antibiotic activities were isolated from microalgae for the first time by Pratt et al., in 1944. They isolated a fatty acid mixture from unctions . The antrculosis . Salvatostrongly .Skeletonema tropicum and Chaetoceros pseudocurvisetus for their ability to perform anti-tuberculosis activities. They reported that anti-tuberculosis activity only presented when these two microalgae were cultured under control and phosphate-limited situations, but not under a nitrogen starvation condition. The organic extracts of Skeletonema tropicum and Chaetoceros pseudocurvisetus performed an anti-tuberculosis activity, but not cytotoxicity against human cells [Nostoc sp. MGL001 and was shown to be toxic to Dalton lymphoma cells with an IC50 of 372.4 ng/mL in a 24-h assay [Lauritano et al. (in 2018) investigated an cells . In addi-h assay .dinoflagellate Amphidinium carterae extract has also been evaluated, and the biologically active compounds were identified. It was found that the extract inhibited Staphylococcus aureus (Gram-positive), Klebsiella pneumoniae, and Escherichia coli (Gram-negative), Mycobacterium tuberculosis, and the fungus, Aspergillus fumigatus [Scenedesmus spinosa and they found compounds that could perform an antibacterial activity. The extracts of water, dimethyl sulfoxide (DMSO), and acetone all inhibited the growth of Bacillus subtilis. Interestingly, the DMSO extract, but not the water or acetone extract, showed inhibitory effects on Klebsiella pneumoniae and Escherichia coli [cyanobacteria Lyngbya majuscula, which performed strong antimicrobial activity with the minimum inhibitory concentration of 6.7 μM [The antibacterial activity of the umigatus . Daniellhia coli . Levert f 6.7 μM .Chlorella sorokiniana, Chlorella sp., and Scenedesmus sp., and tested this ME (methanol extraction) for the presence of activity against bacteria. The ME from Chlorella sp. demonstrated that it performed an antibacterial activity with effective concentrations at 0.312 to 6.25 g/L. The results of the MTT cell viability assay showed that these MEs were not toxic to Vero cells, demonstrating their safety. The major components of the ME from Chlorella were determined using a GC-MS. It contains 18.5% phenol, 18.3% hexadecanoic acid, 14.43% phytol, 13.69% 9,12-octadecadienoic acid, and 7.23% bicyclo heptane [Scenedesmus obliquus and the fractions derived from it were evaluated by Diaa et al., The crude diethyl ether extract had an inhibition zone between 12.5 and 19.5 mm for all the bacteria that were tested and between 8.7 and 18.3 mm for the fungi that were tested. A further purification of the extract using column chromatography identified two fractions that performed high antimicrobial activities against the bacteria and fungi that were tested [Abdult et al. (in 2022) used methanol to extract  heptane . The ante tested .Chaetoceros muelleri in an attempt to find antibacterial activities. The activities of these extracts against bacteria, including Mycobacterium tuberculosis, were tested. The hexane extract performed the strongest antibacterial activity against all the bacterial strains that were tested, showing a potent activity against Mycobacterium tuberculosis. The minimal inhibitory concentration was found to be at 100 μg/mL. On the other hand, the ME was not effective, with the IC50 being greater than 800 µg/mL, whereas the IC50 values of other extracts were between 267 and 142 μg/mL. Interestingly, the ME was the only one that did not show cytotoxicity in the Vero cells [Daniela et al. (in 2022) have used acetone, chloroform, dichloromethane, ethyl acetate, methanol, and hexane to extract the biomolecules from ro cells .in vivo. If more in vivo experiments were performed, the applications of the antibacterial effect of microalgae in health will be realized, increasingly.The main purpose of isolating compounds that can perform antibacterial biological activities from microalgae is to discover new drugs, new food preservatives, or new compounds for agricultural biocontrol applications. Most of the current studies of these antibacterial activities are conducted in vitro rather than Leptolinngbya boryana, Arthrospira platensis, Nostoc punctiforme, Oscillatoria sp., and Leptalynbya sp. on two viruses, Coxsackie virus B3 (CVB3) and rotavirus (RV). The therapeutic indices (TI) of the MEs from Leptolinngbya boryana, Arthrospira platensis, Nostoc punctiforme, Oscillatoria sp., and Leptalynbya sp. were found to be 50.0, 30.0, 27.6, 16.6, and 20.0 for CVB3, respectively. In addition, the TI values of the extracts from Arthrospira platensis and Oscillatoria sp. were at 45 and 42.5 for RV, respectively. On the other hand, the TI values of the extracts from Leptolinngbya boryana, Leptolyngbya sp., and N. punctiforme were from 2.8 to 7 for RV, which are relatively low. The results of this study indicate that the extracts from these five microalgae performed antiviral activities against CVB3 and RV, suggesting the presence of natural and bioactive compounds that are useful for further study [Deyab et al., have investigated the effects of extracts from five cyanobacteria, er study .Porphyra polysaccharide can inhibit the replication of RSV, herpes simplex virus 1 (HSV-1), and CVB3 viruses. The underlying mechanism is to prevent the virus from entering the host cells by interacting with virus particles or host cells [Chlorella can inhibit the replication of HSV-1 and prevent the attachment of viral particles to cell membrane for their entry [st cells . The extir entry .Euglena gracilis Z to mice orally and observed that it significantly reduced the influenza virus titers in the lung and increased the levels of cytokines such as IFN-γ, IL-1β, etc. These results suggest that the antiviral activity of Euglena gracilis Z is performed through immune regulation [Euglena gracilis Z treatment was able to relieve the symptoms in mice that were infected with the influenza virus. It effectively attenuated the infections of all the influenza viral strains that were tested, including those strains that are resistant to Oseltamivir and Amantadine, which are two anti-influenza medications. A further analysis of the mechanisms showed that the Euglena gracilis extract did not suppress viral replication. Either the pretreatment or the extended treatment with the extract was sufficient to reduce the viral titers in the cells that were infected by the influenza virus, suggesting that the target of the Euglena gracilis extract is the host cellular functions, rather than the viral replication per se [Spirulina has been shown to inhibit HSV-1 infection with the same potency as acyclovir. In the in vitro experiments, the Spirulina extract was able to block the interactions between the vital particles and the host cells, which lead to reduced virial adhesion and entry to the cells. In clinical studies using a herpes exacerbation model, creams containing calcium spirulan and microalgae extracts were more effective in terms of prevention than creams with Acyclovir were [Spirulina platensis is very low. Therefore, doses of Spirulina platensis extract of up to 5000 mg/kg are safe and tolerated in model animals. Anti-influenza efficacy studies have shown that the Spirulina platensis extract inhibits the plaque formation of several influenza viral strains, even including the Oseltamivir-resistant strains. The extract of Spirulina platensis functions in the early stages of viral infection and production in the host cells, which improves the survival rates of mice that are infected with influenza and inhibits hemagglutination due to the influenza infection [Nakashima et al. (in 2017) administered gulation . Euglenan per se . The extvir were . The cytnfection .Dunaliella (D. tertiolecta), in vitro. The results showed that the water extracts significantly inhibited the replication of a viral haemorrhagic septicaemia virus and had a trend  to inhibit the replication of the African swine fever virus. The active ingredients in the water extract of D. tertiolecta may be sulfated polysaccharides as polymer sulfated dextran (500 kDa) also has antiviral activity. Therefore, the responsible bioactive molecules in the water extracts of other microalgae might be also sulfated polysaccharides with a molecular mass ranging from hundreds to thousands Da, as experiments have shown that sulfuric acid glucan with masses from 5 kDa to 100 kDa are inactive [Trichodesmium erythraeum. Three of the five, debromo-dolastatin, anhydrodebromo-dolastatin, and 3-methoxy debrominated dolastatin at concentrations of 0.1–10.0 μmol/L, exhibited a dose-dependent inhibitory effect on the chikungunya virus. The IC50 and selectivity indexes of debromo-dolastatin, and 3-methoxy debrominated dolastatin were estimated to be 1.3 and 2.7 μmol/L, and 10.9 and 9.2, respectively [Fabregas et al. (in 1999) investigated the antiviral activities of water extracts of 10 marine microalgae, including inactive . Gupta eectively .The antiviral activities of molecules that were derived from microalgae are related to many factors such as their growth stage, the habitat areas and environment that they are found in, the extract fractions, and the methods of examination. It appears that we can find new antiviral active substances from microalgae. However, their contents are generally very low, which are not sufficient to replace chemical and synthetic antiviral agents. With the progress of microalgae bioengineering technology, the applications of biological cultures, and genetic engineering and other technologies provide a simple and feasible way to obtain large amounts of antiviral substances from microalgae for their clinical uses.Spirulina can be used to reduce skin wrinkles and prevent skin aging that is caused by UVB radiation. It has been speculated that the C-PC treatment facilitates the expression levels of MMP-1 and MMP-9, which participate in the skin tissue remodeling. In HaCaT cells, the C-PC treatment stimulated the expression levels of loricrin, filaggrin, and involucrin, which are the proteins that are protective to UV light damage [C-phycocyanin (C-PC) that has been extracted from t damage ,65.AQP3 was significantly increased in a concentration-dependent way in both cell lines. AQP3 protein expression levels were also increased when astaxanthin was added to HaCaT cells. In the EpiSkin 3-D human epidermis cell culture model, the presence of astaxanthin is also increased the AQP3 expression. Moreover, the addition of astaxanthin facilities the permeability of glycerol in the EpiSkin 3-D model. The permeability of glycerol in the astaxanthin and glycerol group is higher than that in the glycerol-only group. Both the AQP3 mRNA and its protein activity in the skin were elevated after the astaxanthin treatment [The supplementation of astaxanthin has been shown to maintain the structural integrity of the skin, minimize wrinkle formation, and improve water retention in the skin. The use of astaxanthin also blocks the UV-induced damages in the skin, without causing any obvious detrimental effects. In clinical practice, a daily supplementation of 3 to 6 mg astaxanthin is protective to the aging process of the skin that is due to exposure to sun light . When asreatment . Either reatment .Nannochloropsis ethanolic extracts have a variety of skin protection functions. The ethanol extracts contain significant amounts of PUFAs such as EPA, carotenoids such as β-carotenes, astaxanthin, and zeaxanthin, and phenolic compounds. Based on the results of tyrosinase inhibitory activity, the ethanol extracts demonstrated multiple skin protective functions that include anti-melanin, moisturization, antioxidant, anti-inflammation, anti-photoaging, and anti-wrinkle. All these activities can help to restore cell viability after UV exposure [Sun et al. (in 2021) reported that exposure .S. platensis. These peptides included P2 (MAACLR), P3 (RCLNGRL), P4 (RYVTYAVF), P5 (SPSWY), and P7 (AADQRGKDKCARDIGY). The authors tested the inhibitory activities of these peptides against tyrosinase of mushroom and B16-F10 mouse cells, and they reported that P5 and P7 were effective in reducing the tyrosinase activities. P5 was the most potent and had an IC50 of 12.1 µM for the mushroom tyrosinase. In addition to inhibiting tyrosinase, P2 and P3 can scavenge free radicals with an IC50 that is lower than 200 µM. For the tyrosinase of B16-F10 mouse cells, P5 and P7 performed inhibitory activities and had IC50 values at 48.9 and 34.2 µM, respectively. Interestingly, P4 can inhibit mushroom tyrosinase, but promoted melanin synthesis by three times in the B16-F10 cells. The results show that microalgal proteins can be considered as a source for biomolecules that can regulate melanin production [With the help of a computer, Kose A et al. (in 2022) identified several peptides from the digested products of oduction .Chlamydomonas reinhardtii extract on melanin synthesis have been studied in B16F10, a human epidermal cell line, and other models mimicking human skin. The treatment with the Chlamydomonas reinhardtii extract dose-dependently inhibited α-melanocyte-stimulating hormone-induced melanin synthesis. In addition, this extract treatment also suppressed the mRNA and protein levels of tyrosinase, tyrosinase-related protein 1 and 2, and microphthalmia-related transcription factors. These inhibitory effects are associated with the reductions of the activation states of protein kinase A and the other up-stream kinases of melanin production [The effects of the oduction .Scenedesmus rubescens on the biomarkers of UV-irradiation-induced photodamage, such as decreased viability, decreased collagen content, hyperpigmentation, and sunburned cells, in, in vitro and ex vivo skin models. The extracts were shown to reduce the signs of cellular aging that were caused by UV exposure. The extract treatments increased the viability of dermal fibroblasts, rescued the content of dermal collagen, reduced the formation of sunburned cells, and inhibited the tyrosinase activity [HO-1 gene. In addition, it has been found that the phycocyanin treatment induced the nuclear translocation of nuclear factor (erythroid-derived 2)-like 2 (Nrf-2). Furthermore, the phycocyanin treatment attenuated UV-stimulated apoptosis and the expressions of p53 and Bax, and the activation of caspase-3. This treatment led to the phosphorylation of protein kinase C (PKC) α/β II. When PKC α/β II is specifically inhibited by its inhibitor Go6976, the phycocyanin-mediated induction of HO-1 expression is blunted. Furthermore, the small interfering RNA-mediated knockdown of HO-1 stimulated the cleavage of poly (ADP-ribose) polymerase 1, and the activation of caspase-3 after the pretreatment of phycocyanin. All of these results demonstrate that the phycocyanin-stimulated HO-1 expression needs an intact PKC α/β II-Nrf-2/HO-1 pathway, and the phycocyanin treatment attenuates UV-induced apoptosis in skin cells [Researchers have evaluated the effects of extracts from the freshwater microalgae activity . The proin cells . The uses of microalgae extracts have been shown to improve dermal health, which is mediated by the direct inhibition of several key players in the oxidative stress cascade, and inflammation, concurrently. This has been accepted as the main mechanism by which the microalgae extracts protect the skin from damages that are due to the exposure to sun light.Chlorella and Spirulina on obesity have been studied in C57BL/6 mice that were fed a high-fat diet (HFD). The polysaccharides from these two microalgae were equally effective in attenuating body weight gain, disturbance of glucose and lipid metabolisms, inflammation, and a fatty liver in the HFD-fed mice. In addition, the microbiota in the gastrointestinal (GI) tract was also improved with the increase in the relative abundance of beneficial bacteria such as Clostridium, Bacteroides, and mollusks, and a decrease in the harmful ones such as Actinomyces and Verrucobacterium [Nannochloropsis oceanica. The treatment with Nannochloropsis oceanica promoted the retention of lean body mass in these animals, which is attributed to the elevated intake of dietary protein and an increase in the beneficial bacteria such as Oxyphotobacter in the GI tract [The effects of polysaccharides from acterium . du PreeGI tract .Spirulina flatus has been shown to prevent the rapid rise of blood glucose and insulin levels and correct the hepatic enzymes in streptozotocin-induced diabetic mice that are fed an HFD. The Spirulina flatus treatment also improved the plasma lipid profile and inflammatory markers such as adiponectin and TNF-α. The hepatic expression levels of sterol regulatory element-binding protein 1c (SREBP-1c), a master regulator of lipogenesis, is also reduced after the Spirulina flatus treatment, indicating its effect on the fatty liver. In addition, the Spirulina flatus treatment increased the hepatic expression levels of peroxisome proliferator-activated receptor α (PPARα), PPARγ coactivator-1α (PGC-1α), and mitochondrial transcription factor A, which are proteins promoting the mitochondrial biogenesis, indicating the elevation of energy expenditure in the treated animals [The use of  animals .Tisochrysis lutea on obesity and metabolic disturbances in rats. The supplementation of Tisochrysis lutea did not affect plasma alanine aminotransferase, indicating that were was tolerance to this treatment. On the other hand, the treatment reduced body weight, blood levels of glucose and insulin, and increased the levels of total and high-density lipoprotein (HDL-C), but not low-density lipoprotein cholesterol (LDL-C) and triacylglycerol (TAG) levels. The Tisochrysis lutea treatment also reduced the adipose tissue mass, blood leptin and TNF-α levels, and TAG and cholesterol contents in the liver, without any change in the blood IL-6, IL-4, and LPS concentrations. The expression level of IL-10 in the abdominal fat tissue was increased [Sargassum siliquosum treatment and they found that it reduced abdominal fat deposition and liver fat droplet sizes [Mayer et al. (in 2021) investigated the effects of ncreased . Preez eet sizes .Arthrospira significantly reduced fasting blood glucose (FBG), total cholesterol, and TAG levels, and it increased the HDL-C level without any change to the glycated hemoglobin A1c (HbA1C) and LDL-C. On the other hand, other studies have shown that the Arthrospira supplementation decreases FBG and HbA1C levels in animals with diabetes. In humans, the use of Arthrospira at a does that is less than 2 g per day for less than 2 months significantly reduced FBG. It appears that the Arthrospira supplementation is effective to reduce FBG, TAG, total cholesterol, and HDL-C levels in diabetic animal and humans, indicating its antidiabetic hyperlipidemic activities. The discrepancy of the effects of Arthrospira on HbA1C in the humans and animals is an interesting phenomenon, which requires more studies to reveal the underlying mechanisms that are involved in it [Many studies have demonstrated that the dietary supplementation with ed in it .Nannochloropsis gaditana supplementation on glucose metabolism in streptozotocin-induced diabetic rats. The animals were fed a diet with or without supplementation of 10% Nannochloropsis gaditana for 2 months. The results showed that the Nannochloropsis gaditana supplementation reduced their blood glucose and HbA1c levels and attenuated the amount of tissue oxidative stress and inflammation, which resulted in the improvements of functions in the liver and kidney of the diabetic rats [Nacer et al. (in 2020) have studied the effects of tic rats .Chlorella pyenoidose and studied its effects on pancreatic lipase. The synthetic peptide at 200 μg/mL is effective to inhibit the pancreatic lipase activity, which is attributed to its binding to the active site of the enzyme and the formation of hydrogen bonds by docking modeling. Furthermore, in 3T3-L1 cells, the synthetic peptide at 600 μg/mL significantly decreased the intracellular TAG accumulation (27.9%) to the same extent as 10 μM simvastatin (24.1%), the positive control drug. Further analysis of the mechanisms indicate that the peptide inhibits TAG accumulation and lipogenesis in 3T3-L1 adipocytes. The effect may be achieved by the mediation of major players in lipid metabolism such as SREBP-1c and cell energy sensing signaling proteins such as AMP activated protein kinase, suggesting its potential as a medication for obesity treatment [Zhang et al. (in 2019) reported a peptide, LLVVYPWTQR, from reatment .Chlorella zofingiensis and astaxanthin inhibited the formation of N-(1-Carboxymethyl)-L-lysine, which is mediated by the inhibition of intracellular oxidative stress [Chlorella and diatoms have been shown to perform strong anti-advanced glycation end products activity, which may be attributed to astaxanthin and EPA . Scholare stress . In a rae stress .Many of the metabolic diseases require long-term medications that often have certain side effects. Microalgae have therapeutic and preventive potentials and can be developed to become a new type of healthy foods for patients with metabolic diseases.Aurantiochytrium sp. on neural development and functions have been studied. The extract of Aurantiochytrium sp. was able to attenuate the β-amyloid-induced cytotoxicity and increase ATP production via the correction of metabolism in cells. The treatment promoted spheroid formation and increased the proliferation of β-III-tubulin positive neurons and glial fibrillary acidic protein (GFAP)-positive astrocytes. When the Aurantiochytrium sp. extract was orally administrated in the senescence accelerated mouse-prone 8 mice, a mouse model used for studying aging, the treatment reduced the escape latency in the Morris water maze experiment, improved their memory ability, and increased the BrdU incorporation in cells of the hippocampus dentate gyrus region. The effect may be associated with the rapid rises of BrdU + GFAP+ and BrdU + NeuN+, markers of stem cells and mature neurons, respectively [The effects of the microalga ectively .Spirulina contains a plethora of nutrients and other bioactive molecules that are beneficial to brain health. Studies have shown that the Spirulina treatment relieved mental exhaustion and maintained the integrity of cerebrovascular vessel walls in response to endothelial damage and pressure. This helps to prevent and/or alleviate cerebrovascular disease. In addition, the spirulina supplementation appears to improve motor development, communication skills, and cognitive abilities in children experiencing malnutrition [utrition .The fucoxanthin in microalgae also has certain neuroprotective effects. The fucoxanthin treatment not only inhibits the aggregation of β amyloid 1–42 , but also regulates the activations of PI3K, Akt, and ERK-signaling transduction cascades, which prevents the Aβ oligomers-mediated loss of neurons and oxidative stress. The fucoxanthin treatment has been shown to attenuate Aβ1–42-induced neurotoxicity in SH-SY5Y cells . It has Özugur et al. (in 2022) reported that phototrophic microalgae can initiate oxygen production in the brain, which alleviated the oxygen deficit circumstance and rescued neuron viability in the pathological conditions that are associated with hypoxia .DHA as a PUFA has been shown to activate Nrf-2 pathways and protect neurons in rats that are experiencing TBI. The DHA treatment retained neuronal functions, attenuated edema in the brain, and improved the cognitive abilities in the TBI-induced rats. These are associated with the reduction of nitrogenic oxygen species and ROS in the CA1 area and neurons of the hippocampus in the brain. It appears that the DHA treatment promotes the nuclear translocation of Nrf-2, which regulates the protein levels of HO-1 and NAD (P)H: quinone acceptor oxidoreductases 1 in hippocampal regions . Rats wiThe exact mechanism of the neuroprotective activities that are performed by microalgae is not fully understood. However, recent findings suggest that the protective mechanisms may include anti-oxidation, the inhibition of cholinesterase, the protection against β-amyloid aggregation, and the repairment of neuronal damage. The neuroprotective activities that are derived from some microalgal strains are mediated by DHA, carotenoids, polyunsaturated fatty acids, phenolic compounds, and sterols, etc.Microalgae are autotrophic due to them performing photosynthesis and are found in a variety of places . MacroalTherefore, to make full use of the high value-added natural products in microalgae, new functions and resources need to be continuously investigated. It is necessary to explore suitable culture and induction conditions for different microalgae and to produce a large amount of the specific biomolecules. This also includes designing corresponding photoreactors and large-scale culture processors, accordingly. It is anticipated that more and more research will be conducted to fully explore the microalgae as a rich source for bioactive molecules.


Esophageal squamous cell carcinoma (ESCC) accounts for the main esophageal cancer type, which is related to advanced stage and poor survivals. Therefore, novel diagnostic biomarkers are critically needed. In the current research, we aimed to screen novel diagnostic biomarkers based on machine learning. The expression profiles were obtained from GEO datasets  and TCGA datasets. Differentially expressed genes (DEGs) were screened between 47 ESCC and 47 nontumor samples. The LASSO regression model and SVM-RFE analysis were carried out for the identification of potential markers. ROC analysis was carried out to assess discriminatory abilities. The expressions and diagnostic values of the candidates in ESCC were demonstrated in the GSE75241 datasets and TCGA datasets. We also explore the correlations between the critical genes and cancer immune infiltrates using CIBERSORT. In this study, we identified 27 DEGs in ESCC: 5 genes were significantly elevated, and 22 genes were significantly decreased. Based on the results of the SVM-RFE and LASSO regression model, we identified five potential diagnostic biomarkers for ESCC, including GPX3, COL11A1, EREG, MMP1, and MMP12. However, the diagnostic values of only GPX3, MMP1, and MMP12 were confirmed in GSE75241 datasets. Moreover, in TCGA datasets, we further confirmed that GPX3 expression was distinctly decreased in ESCC specimens, while the expression of MMP1 and MMP12 was noticeably increased in ESCC specimens. Immune cell infiltration analysis revealed that the expression of GPX3, MMP1, and MMP12 was associated with several immune, such as T cells CD8, macrophages M2, macrophages M0, and dendritic cells activated. Overall, our findings suggested GPX3, MMP1, and MMP12 as novel diagnostic marker and correlated with immune infiltrates in ESCC patients. Esophageal cancer (EC) is one of the most common malignancies worldwide, which is always accompanied by high morbidity and mortality . EsophagThe investigation of gene expression profiles using microarrays has become a frequent method for locating important hub genes and important pathways . In thisThe microarray dataset GSE20347, GSE38129, and GSE75241 was downloaded from the Gene Expression Omnibus (GEO) database. The GSE20347 dataset included 17 pairs of ESCC and nontumor specimens, whereas the GSE38129 dataset included 30 pairs of ESCC and nontumor specimens. Due to the fact that the GSE38129 datasets share a platform and are important for merging data from a variety of datasets, they have been combined into a metadata cohort for the purpose of doing additional integration analysis. In addition, the combat function contained inside the “SVA” software package in R was utilized in order to eliminate the batch effect. In addition, the validation cohort comprised 15 pairs of ESCC tissues and neighboring normal tissues, which were taken from the GSE75241 datasets.t-tests were utilized. The DEGs are genes that had an adjusted p value that was lower than 0.05 and had an absolute fold change that was higher than 3.The DEG analysis was carried out with the help of the Limma program . In ordep value of less than 0.05 was required.The biological importance of DEGs was investigated using GO term enrichment analysis, which included biological processes, cellular components, and molecular functions. This research was conducted using the “GOstats” program included in Bioconductor. The KEGG pathway enrichment analysis of DEGs was carried out by the “GeneAnswers” Bioconductor program in order to identify important pathways that are closely associated with the beginning and development of ESCC. In order to reach statistical significance and achieve significant enrichment, a When doing five-fold cross-validation, a technique known as least absolute shrinkage and selector operation (LASSO) and support vector machine-recursive feature elimination (SVM-RFE) were employed, respectively, to filter the critical genes , 15. TheIn order to determine whether or not the found biomarkers had any predictive power, we constructed a ROC curve by comparing the levels of mRNA expression in 47 ESCC tumor specimens to 47 nontumor tissues. The value of AUC was used to measure the diagnostic efficiency in distinguishing ESCC specimens from nontumor specimens, and this finding was then verified using the GSE75241 dataset.https://cibersort.stanford.edu/). A mixture sample's relative immune cell fraction was estimated using CIBERSORT and can be applied to compare immune cell populations within and across studies.Based on the reference signature matrix of 547 genes, we employed CIBERSORT to analyze the percentages of various immune cells in tumor and nontumor specimens. When we ran the program with the default LM22 feature matrix at 1000 permutations, we submitted the data of gene expressions generated from the sample mixture file to the CIBERSORT web page . Extensive retrospective analysis was performed on the GSE20347 and GSE38129 GEO datasets, which contained data on 47 ESCC and 47 nontumor samples. We used the Limma package to remove batch effects before analyzing the metadata DEGs. There were a total of 27 DEGs collected: 5 genes were significantly elevated, and 22 genes were significantly decreased .Later, we carried out GO assays using the “clusterProfiler” R package and observed that, in the BP group, the DEGs were mainly involved in extracellular structure organization, skin development, extracellular matrix organization, epidermal cell differentiation, and keratinocyte differentiation. In the MF group, the DEGs were mainly involved in serine hydrolase activity, serine-type peptidase activity, metallopeptidase activity, receptor ligand activity, and extracellular matrix binding . HoweverResearchers made use of two distinct algorithms in their search for possible biomarkers. Regression analysis carried out with the LASSO algorithm helped reduce the number of DEGs, which led to the identification of eight biomarkers for ESCC . The SVMThe diagnostic abilities of GPX3, MMP1, and MMP12 in discriminating ESCC from nontumor specimens confirmed excellent diagnostic values, with an AUC of 0.939 (95% CI 0.879–0.986) in MMP12, AUC of 0.959 (95% CI 0.916–0.990) in MMP1, AUC of 0.985 (95% CI 0.963–0.100) in GPX3, AUC of 0.962 (95% CI 0.922–0.990), and AUC of 0.924 (95% CI 0.853–0.979) . Moreovet-test did not show a distinct difference of GPX3 expressions between ESCC samples and nontumor samples  family belongs to the metzincin superfamily . ExtraceGlutathione peroxidase (GPX) is an important peroxide that has been demonstrated to be widely involved in nontoxic compounds, the reduction of toxic peroxides into hydroxyl compounds, and the decomposing of enzymes , 31. GroThe role of the tumor microenvironment (TME) in the development of the tumor was proven by an increasing amount of evidence . The malHowever, our present study has some limitations. Firstly, considering the limited size of the sample, it will be necessary to do extensive clinical tests. Secondly, we fail to evaluate the expression profile of GPX3, MMP1, and MMP12 in the serum/plasma samples in patients with ESCC. Analyzing the biomarkers present in the serum and plasma samples could be an effective way to evaluate the response to treatment in real time. Additionally, the function of GPX3, MMP1, and MMP12 remained largely unclear, and their function and mechanism were worth further exploration by molecular function experiment. We plan to better incorporate more data sets to demonstrate our findings in the next paper. In order to make these more accurate, we intend to obtain tumor specimens in addition to clinical data and demonstrate the accuracy of the results via tests.We identified GPX3, MMP1, and MMP12 as novel diagnostic genes for ESCC. Our research also provided methods to evaluate those that had a higher potential to benefit from immunotherapy and identified a number of candidate therapeutic targets that could provide a more efficient form of treatments.