Dietary interventions may modulate inflammatory indicators, but the correlations between dietary intervention and inflammatory markers in metabolic syndrome (MetS) settings remain opaque.To evaluate the effects of dietary intervention on interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), and C-reactive protein (CRP) in patients with MetS by systematic review and meta-analysis.Databases, including PubMed, Embase, Cochrane Library, Scopus, and Google scholar, were searched from June 2011 to June 2021 for relevant available articles. Standardized mean difference (SMD) was generated as effect size by meta-analysis for continuous variants, including IL-1β, IL-6, TNF-α, and CRP levels. Then, according to study characteristics by dietary patterns of the intervention, subgroup analyses were performed.p = 0.02, I2 = 64%). However, IL-1β, TNF-α, and CRP levels did not change significantly compared with the control group. Sensitivity analyses further yielded similar results.Finally, 13 studies comprising a total of 1,101 participants were included for the meta-analysis. IL-6 levels in dietary patients were significantly lower than controls (SMD = −0.30, 95% CI = −0.55, 0.04, Dietary intervention may help decrease IL-6 rather than IL-1β, TNF-α, or CRP levels in patients with MetS. Metabolic syndrome (MetS) has become a global epidemic disease due to population aging and lifestyle changes, including diets . The varBad eating habits are the controllable factors accelerating the development of inflammation and related diseases, including MetS . AccordiAlthough some trials have proved that dietary intervention can reduce the serum level of inflammatory factors, it is still controversial if there is an association between dietary intervention and the serum level of inflammatory factors in patients with MetS.Hence, in this study, we aimed to perform a systematic review and meta-analysis to investigate the effects of dietary intervention on IL-1β, IL-6, TNF-α, and CRP levels in MetS.The current systematic review and meta-analysis was implemented in accordance with the principles of the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement .Database, including PubMed, Embase, Cochrane Library, Scopus, and Google scholar, were searched from June 15, 2011 to June 15, 2021. We used the following mesh terms: “inflammatory markers,” “diet” in combination with “metabolic syndrome.”Inclusion criteria were as follows:Randomized controlled trial (RCT) studies;Conducted dietary intervention/s of more than 4 weeks;Meets the diagnostic criteria of MetS;Studies that provided numbers, means, and standard difference (SDs) of IL-1β, IL-6, TNF-α, and CRP.Exclusion criteria were as follows:in vitro study, animal study, or case report;literature reviews, Included patients who had co-morbidities other than MetS; publication data , (2) study design, (3) total number of participants, their age, sex, BMI, and study duration, (4) glucose, insulin, blood lipid levels, and blood pressure status, and (5) mean and standard difference for IL-1β, IL-6, TNF-α, and CRP levels.Qualities of enrolled studies were assessed according to the Cochrane Risk of Bias Tool . Two invp < .05 was considered to be statistically significant. Standardized mean difference (SMD) was generated as effect size by meta-analysis for continuous variants, including IL-1β, IL-6, TNF-α, and CRP levels. If I2 <50% and p > 0.01, a fixed-effect model would be used. Otherwise, a random effect model would be implemented. If I2 > 75%, further analysis encompassing sensitive analysis, subgroup analysis, or meta-regression was carried out to explore the source of heterogeneity. Publication bias was evaluated by funnel plot and Egger's tests.Review Manager 5.3 was implemented in our analyses. Eventually, 13 articles –26 reporIn the included 13 trails, the following six types of dietary intervention patterns were investigated: berry , 20, 26,A risk of bias summary is depicted in Based on data from 2 trials (101 participants) , 21, we p = 0.02, I2 = 64%). Of the 11 included studies , 18–25, As presented in Based on data from 8 trials (458 participants) –22, 26, p = 0.320). The number of studies that analyzed IL-1β, TNF-α, and CRP levels was <10. Hence, it was inadequate to perform a publication bias test.The scatter funnel plot in IL-6 levels appeared symmetrical, indicating the absence of publication bias. Additionally, Egger's test detected no publication bias , including MetS, diabetes mellitus, obesity, and cancer . This waInterleukin-1β could increase insulin resistance and promote apoptosis of β cells in animals . AlthougInterleukin-6 was shown to be a vital mediator of acute phase response with a pleiotropic effect on inflammation during immune response. We found that IL-6 significantly decreased after dietary intervention, especially with fatty acids diet. It may be that dietary intervention can produce immediate changes in MetS, leading to significant changes in IL-6. The JIS and the IDF definitions substantially identify more individuals with MetS than subjects with MetS diagnosed by the NCEP ATP III/ATP III definitions . As for Tumor necrosis factor-α induced phosphorylation of insulin receptor substrate 1 (IRS-1), preventing insulin from binding to the receptor, which consequently led to insulin resistance . In addiLow-calorie diets had been shown to improve the level of inflammatory factors, particularly with advancing age and improving obesity and MetS parameters. However, this was achieved through a significant reduction of total energy intake and supplementing micronutrients . The resIn addition, one of the most commonly used dietary modifications consists of increasing the protein content of the diet. The role of this modification in inflammation was controversial . The DiOTargeting anti-inflammatory therapies had been applied to patients with MetS. In doing so, the IL1-β inhibitor, Canakinumab, increased insulin secretion . In addiFirstly, most eligible studies did not adjust potential confounding factors. Secondly, due to the influence of gender, race, ethnicity, and social factors, our findings should be interpreted in different geographic contexts. Thirdly, the reason for the analysis might be due to the limited time of intervention for the inclusion of RCT, leading to no significant changes in inflammatory marker levels. In addition, individual differences, different methods of intervention, and sample size might also directly affect the analysis results. Lastly, according to different diagnostic criteria, long-term trends may have confounded the results and limited generalizability.Our study suggests that dietary intervention might decrease IL-6, IL-1β, CRP, and TNF-α in MetS. However, different diets might have different protective mechanisms for MetS. More time and more appropriate dietary patterns are needed to improve the inflammatory state of MetS. In addition, more research is needed to clarify the mechanisms underlying the effect of dietary intervention on this population's inflammatory markers. Anti-inflammatory agents combined with dietary intervention may be therapeutically useful in treating and preventing MetS.The original contributions presented in the study are included in the article/ZZ contributed to the study design, wrote the manuscript, reviewed and edited the manuscript. MW, JL, ZZ, HZ, XC, and NW conducted the literature search and performed data extraction and data analysis. MW, JL, and XC did the statistical analyses. MW, JL, XC, XH, QL, and SC contributed to the writing of the manuscript. All authors approved the submitted 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.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.


The adverse effects of PM2.5 on the respiratory and cardiovascular systems have been extensively studied, while its detrimental effects on the central nervous system (CNS), specifically neurodegenerative disorders, are less investigated. Neurodegenerative disorders are characterized by reduced neurogenesis, activated microglia, and neuroinflammation. A variety of studies involving postmortem examinations, epidemiological investigations, animal experiments, and in vitro cell models have shown that PM2.5 exposure results in neuroinflammation, oxidative stress, mitochondrial dysfunction, neuronal apoptosis, and ultimately neurodegenerative disorders, which are strongly associated with the activation of microglia. Microglia are the major innate immune cells of the brain, surveilling and maintaining the homeostasis of CNS. Upon activation by environmental and endogenous insults, such as PM exposure, microglia can enter an overactivated state that is featured by amoeboid morphology, the over-production of reactive oxygen species, and pro-inflammatory mediators. This review summarizes the evidence of microglial activation and oxidative stress and neurodegenerative disorders following PM2.5 exposure. Moreover, the possible mechanisms underlying PM2.5-induced microglial activation and neurodegenerative disorders are discussed. This knowledge provides certain clues for the development of therapies that may slow or halt the progression of neurodegenerative disorders induced by ambient PM.Fine particulate matter (PM PM is the most widespread health threat and has been strongly implicated in diverse diseases . An impom, UFPM) . PM2.5 ey metals ,5. PM2.5n health . Globallannually . Recent ne PM <0. μm, UFPM2.5 has received much attention. Many studies have reported association of PM2.5 exposure with neurological disorders, such as stroke, dementia, Alzheimer’s disease (AD), Parkinson’s disease (PD), and mild cognitive impairment [2.5-induced neurological disorders have not been well elucidated, with oxidative stress and neuroinflammation being two major recognized ones [Since anatomopathological evidence from canine and human residents in Mexico City was reported in the early 2000s ,9,10, thpairment . Thus fazed ones . Due to zed ones , the latzed ones . Oxidatized ones . 2.5 [2.5-induced oxidative stress and neurodegenerative disorders from postmortem examinations, epidemiological investigations, animal experiments, and in vitro studies, with an emphasis on microglial implications in these pathophysiological events. In addition, the potential mechanisms underlying PM2.5-induced microglial activation and associated oxidative stress and neurodegenerative disorders are also discussed. Microglia are the principal players in the brain’s innate immune response . Emergin2.5 ,17,18. T2.5 ,20, but Glial cells account for more than 90% of cells in the human brain and are divided into two populations: the macroglia  and microglia . MicroglUnder resting conditions, microglia survey the microenvironment in real-time with their ramified, motile, fine, and long cellular processes . MeanwhiFrom the above, activated microglia show a broad spectrum of phenotypes ranging from the pro-inflammatory, potentially cytotoxic M1 to the anti-inflammatory, scavenging, and regenerative M2.The first histopathological evidence for a link between air pollution and neuropathology came from a necropsy study with canine residents naturally exposed to a highly polluted environment in Mexico City . In this42, the 42-amino acid form of Aβ, in frontal cortex, hippocampus, and OB [42 and α-synuclein), oxidative stress in frontal and infratentorial neurons and microglia (8-hydroxydeoxyguanosine (8-OHdG) and nitrotyrosine), and frontal BBB impairment, as well as the reduction of the neuroprotective cellular prion protein (PrPC) in the frontal cortex [42, deposition, activated microglia, and reactive glial fibrillary acidic protein (GFAP) positive astrocytes throughout the brainstem [The similar findings were also observed in autopsy examinations of human samples . Adult h, and OB . In contrainstem . Early orainstem . It is nrainstem , APOE4 crainstem ,47. Taken together, the preliminary evidence of postmortem studies indicates that exposure to ambient air pollutants is associated with microglia activation, oxidative stress, and neurodegenerative alterations in brain tissues. However, due to the complex mixture of air pollution, the causative association of air pollution exposure with the observed neurological effects of CNS remains to be specified.There is a growing body of epidemiological studies reporting ambient air pollution-associated neurodegenerative disorders  of Mexico City children [2.5 [Several other epidemiological studies found that exposure to PMk for AD ,54,55,56children , a very children . In addichildren ,59. Givechildren , the agichildren . Thereforen [2.5 .2.5 exposure with the risk for PD. However, the epidemiological results appear inconsistent. For example, one study showed opposite associations between ambient PM2.5 exposure and the incidence of PD among two populations from two locations with different severity of air pollution [2.5 and PD risk in female never-smokers [10 or PM2.5 exposure and the incidence of PD [A few studies have examined the association of ambient PMollution . Another-smokers . Howeverce of PD .Thus far, the epidemiological evidence specifically linking microglial activation to air pollution-induced neurodegenerative disorders is mainly from the studies with children and young adults in Mexico City ,67,68. O2.5-associated microglial activation with neurodegenerative disorders have been investigated among the elderly. In a panel study, Tang et al. examined the air pollutant (including PM) exposure in Chinese people aged 60–69 years, showing that air pollution exposure could induce alterations of neurodegenerative biomarkers, such as Aβ40, Aβ42, α-synuclein, PrPC, Tau (pThr181), and the activation of microglia represented by the over-expression of S100B and microglial triggering receptor expressed on myeloid cells2 (TREM2) [2.5 concentration increments were associated with increases in the biomarkers of neural damage, including a neurofilament light chain (NfL), neuron-specific enolase, and the activation of microglia (S100B) in serum [2.5, such as Cu, Zn, Ni, Mn, Sn, V, Rb, Pb, Al, Be, Cs, Co, Th, U, Cl−, and F− were found to be significantly associated with serum levels of NfL [More recently, the association of PM (TREM2) . Our recin serum . Meanwhis of NfL .2.5 exposure with neurodegenerative disorders.In summary, the available evidence in humans, albeit limited and variable, is suggestive of the association of PM2.5, especially DEP and UFPM, on AD-like pathologies -cleaving enzyme), and COX-2, as well as a decrease in APP, with a minimal change of phosphorylated tau [2.5 for 4 weeks induced a dose-dependent increase in IL-1β and TNF-α in the blood and hippocampus of mice [1.0 for 3 and 6 months resulted in microglia activation in the hippocampus [In AD condition , microgl lesions ,76. The ated tau . The oro of mice . The whopocampus . In addipocampus . 2.5 and are commonly used as a surrogate model of air pollution in health effects studies [3, for 1 month) increased IL-1β, TNF-α, IL-6, MIP-1α (macrophage inflammatory protein-1α), fractalkine, and Iba-1 in most regions of Sprague Dawley (SD) rats, with the midbrain showing the greatest DEP response [3), probably due to the most microglia in the midbrain [1 (250–300 μg/m3) for 3 and 6 months demonstrated that PM1 induced cytotoxicity, lipid peroxidation, microglial activation, and inflammation as well as autophagy and caspase-3 up-regulation in microglia [42 in brain regions [DEP are a major constituent of ambient PM studies . The sho studies ,83. For response . Meanwhiresponse . The susmidbrain . In addiicroglia . Wistar  regions . An inte regions . 3 for 6 h) caused microglial activation, lipid peroxidation, and reduced neurogenesis in all brain regions, particularly in the hippocampus and the OB [0.2 (300 μg/m3) or of neuronal cells to the same nanoPM (1–10 μg/mL) caused an increase in oxidative stress in lipid rafts associated with an increase in Aβ, the latter was inhibited by the antioxidant N-acetyl cysteine, suggesting that oxidative stress was involved in the pro-amyloidogenic effects of air pollution [Nrf2 knockout (Nrf2−/−) mice were subjected to the intranasal instillation of 1 mg/kg PM2.5 for 28 days. Lower levels of antioxidant enzymes, oxidative stress, microglia activation, inflammation, NF-κB activation, and severe nerve injury were detected in the OB of Nrf2−/− mice compared to the OB of WT mice [2.5 exposure-induced oxidative stress and microglia activation was attributed to its metal contents and glutaminase-containing extracellular vehicles (EVs) in the OB [The role of microglia in DEP-induced neurodegenerative disorders has been explored mostly in mouse models. The acute exposure of adult mice to DEP (250–300 mg/md the OB . The blod the OB . The expollution . Interesollution . These rollution  Furtherm WT mice . In addin the OB .3 or higher) for 6 months increased α-synuclein and Aβ42 levels in the midbrain [3) for 3 weeks [In PD condition, α-synuclein is a major component of Lewy bodies, a pathological hallmark of PD . A contrmidbrain . In anot 3 weeks .In summary, animals exposed to PM ambiently or in controlled experiments reveals the same pattern of neurotoxic effects as in humans although the crosstalk among these events need further clarification.In vitro studies provide insights into in-depth cellular and molecular mechanisms by which PM exposure promotes cellular damage and abnormality, linking to neurodegenerative disorders, such as the alterations of cell viability and apoptosis, the dysfunction of mitochondria, the production of ROS, or the release of pro-inflammatory mediators ,17. Thus2.5 displayed the upregulated mRNA of pro-inflammatory cytokines, such as IL-1β and TNF-α [2.5-induced decrease in cell viability, the increase in the intracellular ROS generation, and the NFκB phosphorylation in BV2 cells [2.5 decreased cell survival as a result of neuroinflammation and the production of ROS [2.5 presented elevated apoptosis, IL-1β, and caspase-1 activity, which could be alleviated by the addition of IL-1 receptor antagonists and ROS inhibitors [2.5 has a role in AD pathogenesis, the underlying mechanisms possibly being PM2.5-induced microglial activation, neuroinflammation, increased ROS activity, and even Aβ production [2.5 may activate microglia, since microglia can be activated in vitro by manganese [A mouse microglial cell line (BV2) exposed to concentrated ambient PMnd TNF-α . MoreoveV2 cells . The acun of ROS ,98. In aoduction . Notablyanganese . 2) did not affect the viability of mouse primary cerebellar granule neurons in vitro. However, the death of these neurons was increased two- or three-fold with simultaneous exposure to DEP and microglia [Microglia were first shown to recognize and respond to PM in an in vitro study using DEP . Generalicroglia ), sugges2O2 production from microglia and protected against DEP-induced DA neurotoxicity in midbrain neuron–glia cultures [In the PD condition, primary neuron–glia co-cultures and the HAPI  microglial cell line were pre-treated with DEP (5 μg/mL) followed by LPS (2.5 ng/mL) and synergistically amplified NO production, TNF-α release, and DA neurotoxicity . Pre-trecultures . Anothercultures . Noticeacultures . Thus, icultures ,100,102.Overall, in vitro studies suggest PM-induced oxidative stress and microglial cell mediated inflammatory and/or oxidative responses as potential mechanisms leading to neurotoxicity, and the increased risk of neurodegenerative disease as seen in epidemiological and animal studies. 2.5 has a crucial role in neurodegenerative disorders possible through the activation of microglial cells [2.5 access CNS; (2) the receptors microglia use to relay PM2.5 neurotoxic signals; (3) PM2.5-induced microglial oxidative stress; and (4) the interactions between microglia and neurons. The aforementioned evidence suggests that exposure to PMal cells . HoweverRoute of CNS Effects: Multiple routes for PM2.5 impact on the CNS have been proposed, two of them are predominant  (s (PRRs) , which mMicroglia-associated oxidative stress: ROS are critical components of the pro-inflammatory signaling pathway in microglia [icroglia . Activaticroglia . In micricroglia . Ambienticroglia . Mitochoicroglia . The eleicroglia . Microglia-neuron interactions: The bi-directional communication between microglia and neurons has been recognized to be critical for maintaining a healthy environment in the CNS and also for the chronic development of neuroinflammation [2.5-induced microglial activation and its interaction with neurons are still unclear and warrant further investigation. ammation . The airammation . Thus faammation . With a ammation . In addiammation . Howeverammation . From th2.5-induced neurodegenerative diseases remain largely unknown. The CNS effects are chronic, beginning in childhood, and may take time to accumulate pathology. Specifically, air pollution has been shown to cause neuroinflammation, oxidative stress, cerebral vascular damage, and neurodegenerative pathology, which all involve microglial activation. Evidence from epidemiological and experimental studies suggests that exposure to ambient PM, especially PM2.5, is associated with neurodegenerative disorders. The interpretation of the intracellular and extracellular pathways participating in the generation of oxidative stress in microglia may be important not only for comprehending the pathophysiological basis for neuron damage in neurodegenerative diseases but also for designing effective strategies to mitigate or even prevent PM2.5-induced neural neurodegenerative damage. While epidemiology has linked an increased risk of stroke, AD, and PD with exposure to PM2.5, further epidemiological and mechanistic studies regarding the association between the components of air pollution and the development of CNS diseases are of pressing concern for human health. In summary, air pollution, together with the increasing age of the global population, pose great threats to public health. Thus far, the mechanisms responsible for PM


KRT6, KRT16, serpin-family genes, and SPRR3 confirms the early involvement of hair follicles and the impairment of barrier function in HS lesioned skin. In addition, our results suggest that adipokines could be regarded as biomarkers of HS and metabolic-related disorders. Finally, the RNA-Seq variant calling identified several mutations in HS patients, suggesting potential new HS-related genes associated with the sporadic form of this disease. Overall, this study provides insights into the molecular pathways involved in HS and identifies potential HS-related biomarkers.Hidradenitis suppurativa (HS) is an inflammatory skin condition clinically characterized by recurrent painful deep-seated nodules, abscesses, and sinus tracks in areas bearing apocrine glands, such as axillae, breasts, groins, and buttocks. Despite many recent advances, the pathophysiological landscape of HS still demands further clarification. To elucidate HS pathogenesis, we performed a meta-analysis, set analysis, and a variant calling on selected RNA-Sequencing (RNA-Seq) studies on HS skin. Our findings corroborate the HS triad composed of upregulated inflammation, altered epithelial differentiation, and dysregulated metabolism signaling. Upregulation of specific genes, such as  Hidradenitis suppurativa (HS), also known as acne inversa, is a chronic inflammatory skin disease involving the pilosebaceous unit, with a worldwide prevalence varying from 0.03% to 4% ,6,7. HS HS pathogenesis is still poorly understood, although hereditary factors have demonstrated the ability to increase risks associated with the development of the skin condition. . It has In the -omic era, the next-generation sequencing (NGS) approach made major contributions to the study of HS ,16. HereHomo sapiens organism filter); had matching healthy skin controls; and had raw data (fastq files) available for each sample. Biopsies collected from lesioned skin of HS patients were considered cases, whereas biopsies collected from the skin of patients without HS, or any other skin condition, were accepted as controls.We searched Sequence Read Archive (SRA)  and GeneDESeq2 package objects, where we screened for DEGs [p-values < 0.05 were acknowledged as statistically significant DEGs.for DEGs . Genes wWe integrated the results produced by the independent groups with a meta-analysis using the RankProd package for R software. The package performs a nonparametric approach based on ranks of fold changes (FC) that detect differentially consistently expressed genes (mDEGs) from independent and replicated experiments. Overall, the meta-analysis approach was based on Lee et al. methodology (2019) .p-value < 0.05 were considered significant.We performed a pathway enrichment analysis with the mDEGs. We searched the REACTOME database, using ReactomePA package  for R soWe performed a variant calling sourcing from the aligned RNA-Seq data. Post-processing was applied using Genome Analysis Toolkit v4.1.2.0  to the ap-value < 0.05) differences in the genotype were selected as potential HS genetic markers. The median value of the RNA expression level was calculated for each genotype; large differences between the genotypes were considered statistically significant according to the Wilcoxon test.To infer the connection between genetic variants and HS pathogenesis, we conducted several approaches. Initially we searched for variants in genes previously associated with HS. Then, we sought variants in new potential HS genes. To unravel the new ones, we selected variants based on some filters detailed as follows. Estimates of HS prevalence vary between 0.03% and 4% among different populations ,35. Thert-test for independent samples; not normal and two genotypes: Wilcoxon-Mann-Whitney test; normal and three genotypes: ANOVA; not normal and three genotypes: Kruskal-Wallis test. In the case of three-genotypes tests, we performed a post-hoc paired Wilcoxon test with FDR correction to identify which genotype was responsible for the overall difference).The Shapiro-Wilk test was used to assess if the counts were normally distributed. To uncover if genotypes were associated with differential RNA expression, we used an appropriate test for normally distributed counts based on the number of observed genotypes  resulted in 25 datasets. Among these, seven studies used microarrays; eight used RNA-Seq; six used quantitative real time polymerase chain reaction (PCR); three used single cell (sc) RNA-Seq; and one used target capture. Since we were searching for RNA-Seq studies only, 17 studies were excluded. Out of the eight RNA-Seq studies, six contained analysis of skin biopsies. Of these six studies, two of them did not have matching healthy skin control, and another analyzed familial HS cases only. Therefore, only three studies were included in our meta-analysis and set analysis. Herein, a total of 51 unique HS lesioned samples, and 30 unique healthy skin samples were considered .In the first study, Lowe et al. (SRAdb id: SRP273719) [In the second study, Gudjonsson et al. (SRAdb id: SRP272659)  focused IL-1β to be highly expressed. First, they quantified the expression levels of approximately 30 mediators in HS lesioned skin and compared the results with the expression patterns from these same mediators in healthy skin control and in psoriasis lesioned skin. After finding IL-1β to be upregulated in HS, they compared the results of HS lesioned skin’s RNA-Seq with the transcriptome of various cell types isolated from healthy patients’ biopsies that were exposed to IL-1β and observed transcriptomic overlapping profiles. RT-qPCR and ELISA analyses were also conducted. Our analysis included samples from three HS patients’ lesioned skin biopsy and four healthy individuals skin biopsy not previously exposed with IL-1β.In the third study, Witte-Händel et al. (SRAdb id: SRP168930)  investigDEFB4A/B [TCN1 [MMP1 [S100A7A [MMP3 [TNF-α, IFN-γ, IL-1β, IL-36, IL-17, MZB1, CD19, CD79A, CXCL13, RETN [RARRES2 [PI3 [SERPINB3/4 [SERPINA1 [KRT6/16 [SPRR1/2/3 [THRSP, AWAT2, DCD, UGT3A2, and WIF1, genes associated with energetic metabolism and skin’s barrier function [AQP2/4/5/6/7/8, FOXA1, and ADIPOQ, may impair these pathways.We performed a meta-analysis, which resulted in 6445 annotated genes, and 3628 mDEGs with an FDR < 0.05 and |log2(fold change)| > 1. From these mDEGs, 1699 were upregulated and 1929 were downregulated . BrieflyDEFB4A/B ,41, TCN1N1 [MMP1 , S100A7A[S100A7A , and MMP[RARRES2 , and skiES2 [PI3 , SERPINBRPINB3/4 , SERPINASERPINA1 , KRT6/16[KRT6/16 , and SPRPRR1/2/3  were obsfunction ,54,55,56× 10−14, gene ratio 109/1663), neutrophil degranulation , PD-1 signaling , interferon gamma signaling , signaling by interleukins , complement cascade , antimicrobial peptides , cell surface interactions at the vascular wall , and g-protein coupled receptor (GPCR) ligand binding  appeared enriched.Pathway enrichment analysis of the mDEGs by reactome resulted in 76 statistically significant enriched pathways . We thenIn addition, a pathway enrichment analysis by PANTHER database was conducted. Summarily, pathways with the highest percentage of upregulated mDEGs enclosed inflammation mediated by chemokine and cytokine, integrin and interleukin signaling, T and B cell activation, angiogenesis, and apoptosis . Where, We then performed a set analysis to extract DEGs commonly found in all three selected studies (saDEGs), as well as in both analyses (cDEGs). These analyses can be located in the We performed the variant calling in the HS lesioned samples and controls. The pipeline identified 323,827 unique variants across all samples. Among them, 190,845 variants were annotated as known SNPs and Indels.After filtering, a total of 73,134 genetic variants present in 12,665 genes were found only in patients, and thus selected as candidate variants for HS pathogenesis. Up to 39,709 novel variants not present on dbSNP and Gnomad were found and marked by an asterisk at the end of the information regarding amino acids change and reference gene at the column “AAChange.refGene”. These results are summarized in PSEN), nicastrin (NCSTN), anterior pharynx defective 1 (APH1), and presenilin enhancer 2 (PSENEN)), modulates NOTCH pathway by cleaving and releasing NOTCH’s intracellular domain [NCSTN gene, 3 in the APH1A, 6 in the APH1B, 8 in the PSEN1, 3 in the PSEN2, and 2 in the PSENEN. One variant was already reported in HS patients: a stop gain mutation NM_172341.4: c.168T>G (p.Tyr56X) of the PSENEN gene previously associated with concomitant HS and Dowling Degos Disease (DDD), a rare genodermatosis, classically characterized by acquired reticular hyperpigmentation in flexural sites [MEFV gene, previously associated with HS, has also been spotted [The variant calling analysis identified 34 variants in genes of the γ-secretase complex. This complex, composed of 4 genes , was found by the variant calling, and upregulated expression of important molecules for the IL-1 signaling, as well as genes that stimulate or are stimulated by the expression of this cytokine were found in the mDEGs.Overall, our results were in agreement with the three studies used in our analyses. Lowe et al. , when coiltrates . In normiltrates . Howeveriltrates . Upregul abscess . Indeed, abscess , who alsIFN-γ, but not for IL-1β. Instead, they found upregulation of IL36A and IL36G genes. Still, these genes have been recently reported as agonistic cytokines in the interleukin IL-1 superfamily through the activation of nuclear factor-κB (NF-κB) and mitogen-activated protein kinase (MAPK) [IL-36A and IL-36G were also found upregulated in the meta-analysis. Hessam S. et al. [Gudjonsson et al.  obtainede (MAPK) . IL-36A . et al.  provide . et al. .IL-17A and IL-17F. In HS, the release of follicular debris in the dermis results in the activation of an immune response mediated by Th17 [IL-17, such as DEFB4A/B, MMP1, and MMP9 were also found upregulated, and had the highest-ranking product positions in the meta-analysis. Other skin genes, namely psoriasin (S100A7) and calprotectin (S100A8/9), whose expression augmented in the presence of IL-17, were also found upregulated in the meta-analysis. Since keratinocytes are believed to have an inflammatory profile in HS skin [In our analyses, we found the upregulation of  by Th17 . IL-17 i by Th17 . However HS skin , these fCXCL13, a B cell chemoattractant [MZB1, CD19, CD79A, and CXCL13 were found to be expressed 10 times more in HS lesions when compared to controls. This may suggest that B cell alteration was a common event for HS development. B cells are important for HS pathogenesis since they contribute to fibrosis, stromal remodeling, and therapeutic responses [Enhanced B cell signatures were found by Lowe et al.  and Gudjtractant . In our esponses . The latesponses  in some esponses . When wePI3), an elastase-specific inhibitor that acted as an antimicrobial peptide and was expressed by epithelial and certain immune cells, appeared eight times more expressed. Besides acting against gram-positive and gram-negative bacteria, fungal pathogens, and being involved in NF-κB pathway modulation, cytokine secretion and cell recruitment [SERPINB4, SERPINB3, and SERPINA1 stood out in our ranking product, corroborating the involvement of the hair follicle unit [PI3 and SERPINs may be associated with keratinocyte hyperproliferation responsible for hair follicle plugin, a hallmark of HS.The main enriched pathways associated with skin regulation were ECM organization, keratinization, formation of the cornified envelope, epidermal cell differentiation, and collagen formation and degradation. Moreover, several variants associated with disruption of these pathways were identified, suggesting potential HS-related genes. Zouboulis C.C. et al.  demonstrruitment , the genruitment ,84. Addicle unit . Serpinscle unit . In genecle unit . These rcle unit ,86, two cle unit ,88. ThusKRT6 and KRT16 are particularly interesting since they are expressed together in the outer root sheath of the hair follicles under stressful conditions [KRT6-KRT16 are co-expressed to respond to barrier breach, stimulating hyperproliferation of interfollicular keratinocytes and modulating the inflammatory response to wounds [SPRR3, and several subtypes of SPRR2 and SPRR1 also provided evidence of impairment of barrier functions in HS. SPRRs proteins play a structural role in the cornified envelope, and SPRR3 is generally not detected in normal skin [SPRR3 could impact barrier function through altered production of cornified envelope scaffold that impaired the supramolecular organization of lamellar body-derived lipids into normal bilayer structures [Additionally, several cytokeratin appeared dysregulated, which may affect keratinization and formation of the cornified envelope. nditions . In conto wounds . Dysreguo wounds ,91. In pmal skin . It had ructures ,94. Thisructures . Thus, uMMP1 and MMP3 composed some of the highest positions in the meta-analysis overall ranking, suggesting that skin intrinsic components may represent a critical step in disease progression.Equally important to hair follicle fragility is the family of matrix metalloproteinases (MMPs), a zinc-dependent extracellular protease that break down and remodel the ECM . MMPs plWIF1, AQP5, FOXA1, and DCD were found highly decreased in HS skin by our meta-analysis. Respectively, the genes appeared approximately 10, 2, 6, and 24 times less expressed when compared to healthy skin. In normal conditions, sweat glands’ multipotent progenitor cells contribute to repair and skin re-epithelialization [AQP5, several other aquaporin-related genes such as AQP7, AQP6, AQP8, AQP2, AQP4, AQP7P3, AQP7P1, and AQP4-AS1 were also found down-regulated in the mDEGs. Aquaporins are membrane channel proteins that through the bidirectional transport of water, glycerol, and small solutes across the membrane, serve as critical players of the skin barrier [Finally, sweat glands are believed to regulate epidermal homeostasis and wound repair ,101. Genlization ,101. Thulization , corrobo barrier . Besides barrier . In thisMetabolic dysregulation in HS was detected through several downregulated DEGs and pathways including the metabolism of lipids, triglyceride and fatty acid, and glucocorticoid biosynthesis. Additionally, variants found by the RNA variant calling may be associated with several metabolic-related disorders, including metabolic syndrome (MetS). MetS concomitant with HS had already been described ,106,107.ADIPOQ and susceptibility of HS and metabolic comorbidities. The exploration of adipokines’ role in skin and cellular metabolism is still in an early phase.Body mass index (BMI) data was available for two studies. The data confirmed that most of the patients had a BMI > 25. Obesity is supposed to favor HS skin alteration in a few manners . From thNot all the studies provided clinical information of the patients, such as weight, smoking history, and race, nor general information about lifestyle, comorbidities, and different HS phenotypes. Therefore, clinical information was not taken into consideration in our analysis. Still, independent studies provided similar results, confirming the importance of the DEGs found here for HS pathogenesis. Moreover, whole exome or genome sequencing data from the same samples were not available. Thus, we could not confirm the results found by our variant calling, nor could we confirm them by PCR or sanger sequencing. However, known HS-related variants were identified substantiating our findings. Ultimately, our study design involves re-analyzing previously published transcriptomic data and aggregating evidence of transcriptional changes, thus we were not able to confirm our results on the translational level. Notwithstanding, we hope our results and conclusions stimulate new functional studies that focus on confirming our findings at the protein/tissue level. In general, our results are in compliance with what is reported in the literature.PI3, TNF-α, IL-1β, and IFN-γ suggest that the inflammatory process in HS may be linked with keratinocyte hyperproliferation. Moreover, the upregulation of KRT6, KRT16, serpin-family genes, and SPRR3 alongside downregulation of aquaporin-family genes confirm the involvement of hair follicles and the impairment of barrier function in early phases of HS pathogenesis. Finally, our results demonstrate that HS and metabolic-related syndromes such as MetS and obesity may share transcription profiles and suggest that adipokines may be potential biomarkers for this interaction; down-regulation of ADIPOQ may be associated with diseases’ co-occurrence susceptibility. Several mutations associated with the HS pathogenic triad were found, highlighting potentially new HS-related genes associated with the sporadic form of this disease.Herein, we confirm the HS pathogenic triad composed by upregulated inflammation, altered epithelial differentiation, and dysregulated metabolism signaling. The variant calling and meta-analysis verify this highly interconnected network. The upregulation of


SERPING1 cause quantitative or functional C1-INH defects. Although androgens are not approved for HAE treatment in many countries, they are widely used in China and Brazil to reduce the frequency and severity of HAE attacks. The long-term adverse effects of androgen treatment are concerning for both physicians and patients. Virilization, weight gain, acne, hirsutism, liver damage, headache, myalgia, hematuria, menstrual disorders, diminished libido, arterial hypertension, dyslipidemia, and anxiety/depression are commonly observed during long-term treatment with androgens. These adverse effects can affect the quality of life of HAE patients and often lead to treatment interruption, especially in women and children. In-depth studies of the pathogenesis of HAE have led to the approval of alternative treatment strategies, including plasma-derived C1 inhibitor, recombinant human C1 inhibitor, plasma Kallikrein inhibitor , and bradykinin B2 receptor antagonist (icatibant), some of which have achieved satisfactory results with mostly non-serious side effects. Therefore, a new standard of medical care may expand possibilities for the management of HAE in emerging countries.Hereditary angioedema (HAE) is a rare autosomal dominant genetic disease characterized by repetitive subcutaneous or submucosal angioedema, activation of the kinin system, and increased vascular permeability. C1-inhibitor (C1-INH) deficiency, the main mechanism of HAE pathogenesis, occurs when abnormal activation of plasma kallikrein, bradykinin, and factor XII, or mutation of genes such as  Hereditary angioedema (HAE) is a rare autosomal dominant genetic disease with reported incidence ranging from 1:10,000 to 1:150,000 . PotentiSERPING1) can lead to its quantitative deficiency (type 1 HAE), causing low C1-INH antigen and functional levels, or elicit normal or elevated C1-INH antigen levels, but low C1-INH functionality (type 2 HAE)  has been recommended for short-term and long-term prophylaxis . HoweverLanadelumab, a human monoclonal antibody, is a plasma kallikrein inhibitor approved as a first-line therapy for long-term prophylaxis of HAE attacks. A randomized clinical trial in the United States reported that lanadelumab significantly reduced the attack rate (by 87%) in 27 patients with HAE who received 300 mg of lanadelumab every 2 weeks for at least 6 months. Twenty patients (38.1%) were completely attack-free during the stable state of treatment (days 70–182) . As lanaBerotralstat is an orally administered kallikrein inhibitor that decreases bradykinin production to prevent localized tissue edema during attacks of HAE . In a doFactor XII (FXII) is a key initiator of the kallikrein–kinin system, which produces bradykinin, a central mediator of angioedema, and the inhibition of FXII has become the key area in the discovery of new drugs for HAE. Recently, as a first-in-class, fully human, immunoglobulin G4 monoclonal antibody targeting activated FXII, Garadacimab (200 mg or 600 mg every 4 weeks) could significantly reduce the number of monthly attacks versus placebo and was well tolerated in a randomized, double-blind, placebo-controlled, phase 2 trial study, warranting phase 3 evaluation . In brieAppropriate treatments are essential to improving the QoL of patients with HAE. Because of their wide availability and relatively low cost, androgens and antifibrinolytic agents are the main preventive treatment for long-term prophylaxis of HAE in Brazil and China. In the last decade, several options were also approved for long-term prophylaxis in emerging countries, including C1-INH (via intravenous or subcutaneous administration), lanadelumab, and berotralstat. We also summarized the mechanism of action, half-life, administration route, dosage and regulatory status of androgens, androgen-related drugs, as well as other non-androgenic prophylactic treatments used in HAE up to now. Of note, use of C1-INH is limited by the frequent dosing regimens required for intravenously administered drugs (every 3–4 days). However, subcutaneously administered pdC1-INH concentrate and lanadelumab represent significant recent advances in treatment effectiveness and improved QoL.


Pregestational type 1 (T1DM) and type 2 (T2DM) diabetes mellitus and gestational diabetes mellitus (GDM) are associated with increased rates of adverse maternal and neonatal outcomes. Adverse outcomes are more common in women with pregestational diabetes compared to GDM; although, conflicting results have been reported. This systematic review aims to summarise and synthesise studies that have compared adverse pregnancy outcomes in pregnancies complicated by pregestational diabetes and GDM. Three databases, Pubmed, EBSCOhost and Scopus were searched to identify studies that compared adverse outcomes in pregnancies complicated by pregestational T1DM and T2DM, and GDM. A total of 20 studies met the inclusion criteria and are included in this systematic review. Thirteen pregnancy outcomes including caesarean section, preterm birth, congenital anomalies, pre-eclampsia, neonatal hypoglycaemia, macrosomia, neonatal intensive care unit admission, stillbirth, Apgar score, large for gestational age, induction of labour, respiratory distress syndrome and miscarriages were compared. Findings from this review confirm that pregestational diabetes is associated with more frequent pregnancy complications than GDM. Taken together, this review highlights the risks posed by all types of maternal diabetes and the need to improve care and educate women on the importance of maintaining optimal glycaemic control to mitigate these risks. Globally, it is estimated that 21.1 million (16.7%) live births in 2021 were associated with maternal diabetes ,4. ObesiMaternal diabetes is associated with pregnancy complications and increased rates of adverse maternal and neonatal outcomes ,7. ShortIt is widely reported that all types of maternal diabetes are associated with pregnancy complications; although, adverse outcomes are more common in women with pregestational diabetes ,16,17,18Despite the large body of evidence that associates pregestational diabetes with more frequent adverse pregnancy outcomes than GDM ,23,24,25This systematic review was conducted adhering to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines  Supplem.Three databases, Pubmed, Scopus and EBSCOhost were searched for studies reporting on maternal diabetes and pregnancy outcomes, published between January 1993 and December 2021. The search terms included “type 1 diabetes mellitus” or “type 1 diabetes” or “diabetes mellitus type 1” or “diabetes type 1” and “type 2 diabetes” or “type 2 diabetes mellitus” and “pre-gestational diabetes” or “gestational diabetes” or “diabetes in pregnancy” and “pregnancy complications” or “perinatal outcomes” or “adverse outcomes” or “pregnancy outcomes” and were adapted to each database. An experienced information scientist was consulted to ensure that the search terms were relevant and optimally arranged. References were managed in Zotero 5.0.96.2 . After the removal of duplicate studies, two reviewers (NM and MM) independently screened articles for eligibility. Disagreements or uncertainties were resolved by discussion and consensus or in consultation with a third reviewer (CP). Additionally, references from selected articles were screened for potentially relevant articles.Is there an association between maternal diabetes type and the frequency of adverse pregnancy outcomes?Studies that compared pregnancy outcomes in one or two types of maternal diabetes only, those focusing on other forms of diabetes , etc.), abstracts, review articles, letters, case reports, intervention studies and those not written in English, were excluded. Review articles were screened to identify eligible studies that may have been missed using our search strategy. Studies reporting on adverse outcomes in pregnancies complicated by T1DM, T2DM and GDM were included. This systematic review was conducted to answer the following question:Participants—Pregnant women with GDM;Intervention—No intervention was used in this study;Comparator—Pregnant women with pregestational T1DM and T2DM;Outcome—Pregnancy outcomes.This was achieved using the following:Data that were extracted and recorded included author details (name and date of publication), study details , sample size, characteristics of the population (ethnicity), country and pregnancy outcomes in the different diabetic groups. Two reviewers (NM and MM) independently appraised the study quality and risk of bias using the Newcastle–Ottawa Scale. The Newcastle–Ottawa Scale is used to assess the quality of non-randomized studies, such as case-control and cohort studies . It asseCaesarean section refers to the delivery of a foetus through an incision in the abdominal wall and uterus . PretermA total of 2164 studies were identified from the search strategy. An additional three articles were identified by reviewing the reference lists of relevant articles and reviews resulting in 2167 articles. After removing duplicates, 1958 article titles and abstracts were screened for eligible full-text articles. We excluded studies that compared one or two types of maternal diabetes only, interventional studies, those not written in English, review articles, letters, case reports and abstracts. A total of 20 studies, published between January 1993 and December 2021, met the inclusion criteria and are discussed in this review .n = 196,232 participants; Twenty articles published between 1993 and 2021 were included in the review (n = 2), American Diabetes Association , National Diabetes Data Group , O’Sullivan and Mahan (n = 1), Spanish Group for Diabetes (n = 1), Australasian Diabetes in Pregnancy Society  and World Health Organization 1998/1999 (n = 2). Five studies used institution-based diagnostic criteria, while three studies did not report which diagnostic criteria were used. Pregestational diabetes was determined through hospital records and/or by the medication taken by patients. The studies were conducted in different populations, which included: Omani, Saudi, African, Non-Hispanic black, Australian, Asian, Middle Eastern, Indian, Caucasian and Hispanic. Many of the studies were retrospective and did not report the time of assessment of pregnancy outcomes. Twelve studies included in this review defined one or more of the adverse outcomes; however, definitions and/or cut-offs varied across studies, while eight studies did not define outcomes.The studies in this review used different diagnostic criteria for GDM, which included the International Association of Diabetes and Pregnancy Study Groups Consensus Panel, 2010  of the studies that reported on congenital anomalies reported the overall incidence and not the incidence of the individual congenital anomalies in their comparisons. Due to significant heterogeneity between studies and the low-quality assessment scores for a few studies, a meta-analysis was not performed, as this may lead to an inaccurate estimate of overall effect size .The quality of the 20 studies included in this review ranged from unsatisfactory to very good with scores ranging from 4 to 7 and an average score of 5.5. Three studies scored unsatisfactory (4), seven studies scored fair (5), six studies scored good (6), and four studies scored very good (7) . The stun = 7), followed by preterm birth (n = 7), congenital anomalies (n = 7), pre-eclampsia (n = 6), neonatal hypoglycaemia (n = 5), macrosomia (n = 4), NICU admission (n = 4), stillbirth (n = 4), Apgar score (n = 4), LGA (n = 3) RDS (n = 3) and IOL (n = 2). Of the eleven studies that separately compared pregestational T1DM and T2DM with GDM, the most common adverse outcome reported was C/S (n = 10), followed by preterm birth (n = 7), macrosomia (n = 7), congenital anomalies (n = 6), pre-eclampsia (n = 4), stillbirth (n = 4), neonatal hypoglycaemia (n = 3), IOL (n = 3), Apgar score (n = 3), LGA (n = 3), miscarriage (n = 2), NICU (n = 2) and RDS (n = 1). Certain studies subdivided GDM into true GDM  2 h < 11.1 mmol/L) and overt GDM (fasting glucose ≥ 7 mmol/L or OGTT 2 h ≥ 11.1 mmol/L). For the purpose of this review, we focused on outcomes for true GDM.Of the nine studies that compared combined data for pregestational T1DM and T2DM combined with GDM, the most common adverse outcome reported was C/S . Of the studies that compared pregestational diabetes (combined T1DM and T2DM) with GDM, four studies reported higher rates of C/S in pregestational diabetes compared to GDM [d to GDM ,27,43,46d to GDM ,44. Hyard to GDM . Of the d to GDM ,50,51,52d to GDM . Petticcd to GDM . Soepneld to GDM . In contd to GDM . Taken tPreterm birth. Of the studies that compared pregestational diabetes (combined T1DM and T2DM) with GDM, five studies reported higher rates of preterm birth in pregestational diabetes compared to GDM [d to GDM ,44,45,46d to GDM ,27. Of td to GDM ,50,51,52d to GDM . These rCongenital anomalies. Higher rates of congenital anomalies were reported in pregestational diabetes (combined T1DM and T2DM) compared to GDM in four studies [ studies ,22,25,27 studies . In cont studies ,45. When studies ,17,24,47 studies ,47. In c studies ,21. AlthPre-eclampsia. Higher rates of pre-eclampsia were reported in pregestational diabetes (combined T1DM and T2DM) compared to GDM in three studies [ studies ,45,46, w studies ,27. El M studies . Pre-ecl studies ,24,50. S studies . Taken tNeonatal hypoglycaemia. Three studies reported higher rates of neonatal hypoglycaemia in pregestational diabetes (combined T1DM and T2DM) compared to GDM [d to GDM ,43,46, wd to GDM ,44. Whend to GDM  and Huddd to GDM . Howeverd to GDM . These rMacrosomia. Higher rates of macrosomia were reported in pregestational diabetes (combined T1DM and T2DM) compared to GDM in three studies) [studies) ,22,44, wstudies) . Macrosostudies) ,50. Petistudies) , while Vstudies) . Howeverstudies) ,21,48. ANICU admissions. When NICU admissions were compared between pregestational diabetes (combined T1DM and T2DM) and GDM, four studies reported higher rates of NICU admissions in pregestational diabetes compared to GDM [d to GDM ,43,45,46d to GDM , while Ad to GDM . These rStillbirth. When stillbirth was compared between pregestational diabetes (combined T1DM and T2DM) and GDM, higher rates of stillbirth were reported in pregestational diabetes compared to GDM in two studies [ studies ,46. Howe studies ,45. When studies ,24,49, w studies . AltogetApgar score. Low Apgar scores (<7) were compared between pregestational diabetes (combined T1DM and T2DM) and GDM. Barakat et al., 2010, reported higher rates of low Apgar scores in pregestational diabetes (24.1%) compared to GDM (22.1%) [ (22.1%) ,44,46. L (22.1%) , while tdiabetes .1% compa (22.1%) . These fLarge for gestational age (LGA). Two studies reported higher rates of LGA in GDM compared to pregestational diabetes (combined T1DM and T2DM) [nd T2DM) ,27, whilnd T2DM) . LGA wasnd T2DM) ,52. In cnd T2DM) . AltogetInduction of labour (IOL). Two studies reported no difference in the rates of IOL between pregestational diabetes and GDM [ and GDM ,46. In t and GDM , while P and GDM . In cont and GDM . These rRespiratory distress syndrome (RDS). When comparing pregestational diabetes (combined T1DM and T2DM) and GDM, higher rates of RDS were reported in pregestational diabetes compared to GDM in two studies [ studies ,43, whil studies . In the  studies . These rMiscarriage. When comparing T1DM and T2DM separately with GDM, higher rates of miscarriage were reported in T1DM compared to T2DM and GDM in two studies [ studies ,17. ThesAdverse outcomes associated with maternal diabetes are reported to be more common in women with pregestational diabetes compared to GDM; although, conflicting results have been reported ,27,47,48Thirteen perinatal complications, C/S, preterm birth, congenital anomalies, pre-eclampsia, neonatal hypoglycaemia, macrosomia, NICU admission, stillbirth, Apgar score, LGA, IOL, RDS and miscarriage, which are amongst the most common maternal and foetal adverse outcomes reported in the literature, were compared in this review. C/S was the most common adverse outcome reported. Although it is accepted that not all C/S may be considered an adverse pregnancy outcome , it is oMacrosomia refers to giving birth to babies weighing more than 4 kg and is considered the most common adverse outcome associated with maternal diabetes ,29. The Bax, (Bcl-2-associated X), which is a death-promoting protein associated with increased apoptotic morphological changes and is reversed by insulin [Placental abnormalities and congenital malformations are major risk factors for stillbirth and neonatal death, which represent the extreme end of the spectrum of complications in diabetic pregnancies . Stillbi insulin . In wome insulin . A Cochr insulin .Furthermore, poor glucose control in the third trimester may lead to perinatal asphyxia and low Apgar scores ,76. ApgaLimitations of the studies included in this review may hinder our ability to draw significant conclusions. There was heterogeneity across studies in terms of population characteristics, the diagnostic criteria used, the definitions used for pregnancy outcomes  and different medication regimens . It has been widely reported that ethnicity ,80, advaDespite the inconclusive results from this review, it is evident that pregestational diabetes poses a greater risk for pregnancy complications than GDM and emphasises the importance of maintaining optimal glucose control during the preconception period. Maternal metabolic factors may program physiological adaptation to pregnancy, thereby affecting pregnancy outcomes ,86. The While acknowledging the importance of preconception health and optimal glucose control during pregnancy, the importance of GDM prevention should not be underestimated. As with pregestational diabetes, albeit less common, GDM was also associated with several adverse pregnancy outcomes. Importantly, these complications can be avoided by preventing the development of GDM. During pregnancy, lifestyle modifications that include diet and physical activity have been shown to prevent GDM ,91,92,93The majority of studies included in this review were retrospective. In addition, we did not identify articles that investigated long-term adverse outcomes in women with pregestational T1DM and T2DM, and GDM. Therefore, there is a need for prospective, longitudinal studies in the future to more accurately compare short- and long-term adverse pregnancy outcomes across diabetes types. Preterm birth was one of the most common adverse outcomes reported in this review. The optimal timing of delivery for women with pregestational diabetes is not known due to a lack of published trials ; therefoIn conclusion, the findings from this review confirm that adverse pregnancy outcomes are more common in women with pregestational diabetes compared to women with GDM. These findings highlight the importance of preconception health and the need to educate women of reproductive age who have diabetes or who are at risk of diabetes about the importance of pre-pregnancy care and maintaining good glycaemic control to improve pregnancy health and reduce the risk of adverse pregnancy outcomes. Another important finding of the review is the high rates of adverse outcomes observed in women with GDM, and the need for intervention strategies to prevent the development of GDM.


The detection of circulating tumor cells (CTCs) is an emerging strategy for the early detection, prognostication, and identification of recurrent cancer. The clinical utility of CTC detection has been established, but few studies have employed this strategy for the detection of gynecologic cancers. Here, we present a novel, biochip-based microfluidic device for the detection of CTCs in gynecologic cancers. The study cohort included three patients with cervical cancer, eight with endometrial cancer, two with ovarian cancer, two with breast cancer, and one with vaginal small cell carcinoma. Four cancer type-specific molecular markers , as well as CD13, were used for prognostication and recurrence detection, along with downstream genomic analysis. GATA3 and HER2 were markedly expressed in the patients with cervical cancer, and this expression was strongly correlated with the early detection of recurrent disease. All four molecular markers were expressed preoperatively in the patients with endometrial cancer, and the re-expression of different markers was observed at follow-up before recurrence was confirmed. CD13 was identified as an alternative prognostic marker for both cervical and endometrial cancer. Our pilot study indicated that the novel CTC detection system can be used for prognostication and early detection of disease recurrence, which needed further investigation. Cancer is the most prevalent cause of global mortality, with the majority of deaths attributed to metastatic disease . A surveCirculating tumor cells (CTCs) are cells from the original tumor that enter the bloodstream, where they interact with the microenvironment and ultimately extravasate, resulting in metastasis. The detection of CTCs might serve as a tool for improving disease management by enabling the early detection of cancer, prediction of treatment outcomes, and early detection of recurrence ,4,5,6,7.The detection of CTCs relies mainly on the detection of epithelial surface markers, namely EpCAM. Currently, CELL SEARCH is the only FDA-approved kit on the market. Several studies have reported the clinical utility of CTC detection, with initial success observed in breast, colorectal, and prostate cancer. However, few articles have discussed the utility of CTC detection in gynecologic cancers ,13,14,15The current CTC detection technologies are mainly divided into magnetic beads, microfluidic, and size-based. The magnetic bead method is based on the Cellsearch platform as the mainstream of research, but studies have questioned its low capture rate of CTCs. The method of cell size screening carries the risk that small size CTCs cannot be captured. Microfluidic technology has the advantages of high capture rate and easy operation, and it can locate the cell position with AI image recognition software, which can pick up CTCs accurately and provide high-purity target cell biological information for subsequent molecular analysis.Advances in microfluidic device technology have enabled the detection of rare CTCs more efficently and the identification of several cancer type-specific markers; additionally, epithelial–mesenchymal transition surface markers provide an in-depth evaluation of cancer. Here, we describe the application of a novel method for the detection of CTCs in gynecologic cancer cases and demonstrate its utility in clinical management.A total of seventeen patients, including three with breast cancer, three with cervical cancer, eight with endometrial cancer, two with ovarian cancer, and one with vaginal cancer, were included in our study. Patient demographic characteristics are listed in The three cervical cancer patients included one stage IIIC1 endocervical adenocarcinoma patient (P4), one stage IIIC1 squamous cell carcinoma patient (P18), and one stage IIB squamous cell carcinoma patient (P19). These patients exhibited significant expression of HER2 markers, compared to healthy cohorts . The marPatient 18 (P18) was initially diagnosed with stage IIIC1 squamous cell carcinoma, with persistent elevated serum SCC after CCRT. She underwent adjuvant hysterectomy and bilateral salpingo-oophorectomy, and no histological evidence of disease recurrence was observed. The patient was noted to have increased expression of CD13 at the first follow-up and mild expression of GATA3/PanCK at the second follow-up . A PET sP19, diagnosed with stage IIB squamous cell carcinoma, exhibited initial expression of GATA3, PanCK, and HER2. She underwent CCRT, with no evidence of disease at the time of follow-up. However, all three markers, which exhibited marked expression at the first follow-up, indicated no evidence of disease at the time of writing. Marked expression of CD13 was noted, which was normalized by the first follow-up and increased again by the second follow-up .The eight endometrial cancer patients included three stage IA patients , one stage IB patient (P14), one stage IIIA patient (P10), one stage C1 patient (P13), one stage IIIC2 patient (P11), and one stage IVB patient (P7). All patients exhibited endometrioid histology, except for the carcinosarcoma patient (P13), and all exhibited preoperative overexpression of PanCK, GATA3, HE4, and HER2 . The patP13, diagnosed with stage IIIC1 carcinosarcoma, exhibited initial marked expression of CD13 that persisted throughout the first, second, and third follow-ups. Expression of GATA3/PanCK  was obseP10, diagnosed with stage IIIa endometroid carcinoma, exhibited peritoneal recurrence at the third follow-up, with re-expression of PanCK and GATA3. Initial expression of CD13 was observed. CD13 expression persisted throughout the first and second follow-ups .P5, diagnosed with stage 1, grade 1 endometroid carcinoma, exhibited preoperative expression of all four molecular markers. At the third follow-up, this patient exhibited re-expression of all markers except HER2, with no clinical evidence of disease at the time of writing. CD13 expression fluctuated; increased expression was observed at the first and fourth follow-ups .P3, diagnosed with stage IA, grade 1 endometroid carcinoma, exhibited GATA3/HER2/PanCK expression at the fourth follow-up and CD13 expression at the second follow-up . This paIn all patients that exhibited initial HE4 expression, HE4 expression was undetectable after the first or second follow-up .The two ovarian cancer patients included P1, diagnosed with stage IA immature teratoma, and P2, diagnosed with stage IA mucinous cytadenocarcinoma. Both patients underwent primary surgical intervention, and both exhibited preoperative HE4 expression . No CTCsAll three breast cancer patients had AJCC stage II breast cancer. P8, who had triple negative breast cancer, exhibited marked expression of HER2 and PanCK. P15 and P16 exhibited moderate expression of HER2 . GATA3 rThe patient with vaginal small cell carcinoma exhibited marked expression of GATA3, HER2, and PanCK at the first follow-up , which cA genomic mutation in TP53 was observed in P10, P11, and P14. A mutation in CDH1 was observed in P3 and P5. Two breast cancer patients (P8 and P16) had the BRCA1 mutation .All seventeen patients in our study cohort expressed cancer-specific molecular markers in CTCs, which were detected using V-Biochip microfluidic device technology in conjunction with an automated platform. The importance of identifying recurrence even before clinical evidence of disease was clearly demonstrated in both cervical and endometrial cancer in this study. CD1, an epithelial mesenchymal transition marker, along with the epithelial marker EpCAM, helped to facilitate the detection, monitoring, and prognostication of gynecologic cancer.The prognostic role of CTCs in two of the cervical cancer patients was clearly demonstrated in this small series study. These patients exhibited marked expression of HER2 at the fourth follow-up, which led to the confirmation of a metastatic lesion in P4 . P18 (squamous cell carcinoma) demonstrated persistent expression of CD13 and mild expression of GATA3/PanCK at the second follow-up, which corresponded to avid uptake in the neck lymph node and histological confirmation of metastasis. P19 exhibited marked expression of GATA3/PanCK/HER2 and re-expression of CD13 at follow-up but demonstrated no evidence of disease. Cancer type-specific molecular expression was observed in these three cases, with marked expression of HER2 in endocervical adenocarcinoma and GATA3 in squamous cell carcinoma. All of the confirmed recurrent cases exhibited elevated expression of CD13 at follow-up. Therefore, monitoring CD13 expression in addition to the expression of tumor-specific molecules may be beneficial ,11,13.The eight endometrial cancer cases included one carcinosarcoma case and one dedifferentiated case. Both of these cases exhibited marked expression of GATA3/PanCk and CD13 at follow-up, and recurrent disease was confirmed. The re-expression of markers in P3 and P5 warranted further evaluation. The presence of the genomic mutation in CDH1 indicates that the evaluation of hereditary diffuse gastric cancer and lobular breast cancer should be considered when managing follow-up ,9,10,12.Single-cell genomics and transcriptomics are fields in which further research is necessary ,4,5,6,7.Cells with PanCK expression were detected in some donors in the healthy group. It is possible that normal endometrial cells with PanCK expression  entered Microfluidic technology has the advantages of high capture rate and easy operation ,20,21,22The limitations of this study included the small number of patients and the short follow-up period, making further investiagion into the generalizability of its utility in gynecologic cancer necessary. Studies involving larger cohorts that include patients with additional heterogeneous diseases are needed.In conclusion, our novel CTC detection system, based on microfluidic device technology and an automated platform, enabled detection of the expression of specific epithelial and mesenchymal markers in CTCs. This technology can be used for prognostication and early detection of disease recurrence.From 21 April 2021 to 23 May 2022, seventeen patients  were enrolled in our study. The first follow-up was arranged at 3 months post-operation or 15 weeks after CCRT/RT/CT. The second, third, and fourth follow-ups were conducted at 3-month intervals. . IRB appTM  platform, which is a fully automated platform that uses the V-BioChip microfluidic device for the enrichment and staining of circulating rare cells. The core technology, V-BioChip, is a silicon-based chip produced by metal-assisted chemical etching. The protruding nanostructures of the V-BioChip device are regularly arranged and function in cooperation with the interspace region. The chip undergoes surface treatment, which includes silane-PEG-Biotin deposition and covalent conjugation of streptavidin. This design increases the contact area with target cells. As a result, the target cells can attach to the outer portions of the protruding nanostructures without sustaining punctures or scratches.Instead of the traditional magnetic bead system used for the retrieval of CTCs, this study employed the Cell RevealThe workflow of CTC detection is divided into several steps: collection and preprocessing of the blood sample, enrichment and immunofluorescence staining of CTCs, and scanning and identification of CTCs. Subsequently, target CTCs are isolated with high purity using an automatic cell picker for single-cell whole genome amplification. TM density gradient medium  for the enrichment of the peripheral blood mononuclear cell (PBMC) fraction.For sample collection, 18 mL of peripheral blood was obtained from the patients. The first 2 mL was collected in BD vacutainer K2 EDTA tubes, and the remaining 16 mL was collected in two BD vacutainer ACD tubes (8 mL per tube). Whole blood samples were purified using Lymphoprepg for 5 min to collect the cell pellets and remove the supernatant.Isolated PBMCs were fixed with 4% paraformaldehyde for 15 min at room temperature. Fixed PBMCs were then treated with an antibody cocktail containing biotinylated anti-EpCAM antibody  and biotinylated anti-E-cadherin antibody  and mixed consistently for 30 min at 37 °C. Then, 3 mL of Dulbecco’s phosphate-buffered saline was added to the mixture, which was centrifuged at 400× TM  was used for the enrichment and staining of CTCs. After placing the required reagents in the machine and setting the experimental conditions, the prepared blood sample was injected into the instrument, and the entire process proceeded automatically. The input blood sample was fixed in 4% paraformaldehyde and mixed with 0.1% Triton X-100  and 2% bovine serum albumin to increase the cellular permeability. Subsequently, the sample passed through the V-BioChip device at a flow rate of 0.6 mL/h, allowing the target cells to be captured by the chip.Cell RevealThe CTC target cells were identified using four different antibody cocktails, including CD13/EPCAM, HE4/EPCAM, Her2/Gata3/PanCK, and PAX8/EPCAM. Nuclei were stained with 4,6-diamidino-2-phenylindole .After the completion of cell staining, the V-BioChip was moved to a fluorescence microscope, which was controlled by an automated scanning system  for whole chip image acquisition. The Cell Analysis Tools  system is a tool for cell identification based on image recognition of immunofluorescence staining. The CAT system can screen an entire image within 10 min, identify the target cells, and record the exact position of the target cells on the chip.The target cells were isolated using Cell Picker  and then dispensed into a PCR tube for whole genome amplification. Cell Picker is a system that integrates a motorized upright fluorescence microscope and a micropipette module . The Cel® Spin Miniprep kit . The concentration and purity of the purified DNA were determined using Nanodrop 2000 , and the size distribution was measured using the Agilent 4200 TapeStation with the Genomic DNA ScreenTape assay .DNA from the CTCs was amplified using the PicoPLEX Single-Cell WGA Kit . During each whole genome amplification assay, positive control DNA and a no-template control were used to monitor the amplification efficiency and contamination. After whole genome amplification, the DNA was purified using the QIAPrepTargeted sequencing was performed using the SureSelect Cancer All-In-One Solid Tumor Assay , which includes the following 98 cancer genes: ABL1, AKT1, ALK, APC, AR, ARAF, ARID1A, ATM, BCL2, BCR, BRAF, BRCA1, BRCA2, CCND1, CCND2, CCNE1, CD274, CDH1, CDK4, CDK6, CDKN2A, CDKN2B, CIC, CSF1R, CTNNB1, DDR2, DNMT3A, EGFR, ERBB2, ERBB3, ERBB4, ESR1, ETV1, ETV4, ETV6, EZH2, FBXW7, FGR1, FGR2, FGR3, FGR4, FOXL2, GNA11, GNAQ, GNAS, HNF1A, HRAS, IDH1, IDH2, JAK2, JAK3, KDR, KIT, KMT2A, KRAS, MAP2K1, MAP2K2, MAP2K4, MDM2, MET, MLH1, MSH2, MSH6, MTOR, MYC, MYCN, MYD88, NF1, NF2, NFE2L2, NOTCH1, NRAS, NTRK1, PDGFRA, PDGFRB, PIK3CA, PIK3R1, PTCH1, PTEN, PTPN11, RAF1, RB1, RET, RIT1, ROS1, SMD4, SMARCB1, SMO, SRC, STK11, TERT, TMPRSS2, TP53, TSC1, TSC2, VEGFA, VHL, and WT1.The target CTCs were isolated using the Cell Picker  and then dispensed into a PCR tube containing Tris-EDTA buffer for whole genome amplification. After whole genome amplification, the DNA sample was analyzed using the SureSelect Cancer All-In-One Solid Tumor . The enriched DNA was subjected to next-generation sequencing using Illumina NovaSeq 6000 in a 2 × 150 bp format. The average coverage depth of the captured region was 1000× for CTCs and 50× for germline controls.p-value of <0.05 was considered statistically significant. All statistical analyses were performed using R (R Core Team (2019). R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. https://www.R-project.org/ (accessed on 5 July 2019)).Descriptive statistics were used to analyze the demographic variables. Continuous variables were expressed as the mean and range, and categorical variables were expressed numerically as percentages. A


Global warming and events of heat waves even in temperate climatic zones signify the importance of the genetic makeup of heat stress. Our earlier acute heat stress exposure study (from 30 min to 120 min treatments) in rats led to the designation of aberrant differentially expressed genes (DEGs) and pathways through RNA-sequencing of the three most relevant tissues, i.e., blood, liver, and adrenal glands. However, the causes and mechanisms of differential expression of genes associated with heat stress are still unclear. Using the same RNA-sequencing data, this study identified the differentially expressed mRNA isoforms and narrowed down the most reliable DEG markers and molecular pathways that underlie the thermoregulatory mechanisms. The spatial-temporal differential expression pattern of heat stress-responsive differential mRNA isoforms may be ultimately used in marker-assisted selection for improved thermotolerance.Dnaja1, LOC680121, Chordc1, AABR07011951.1, Hsp90aa1, Hspa1b, Cdkn1a, Hmox1, Bag3, and Dnaja4 were commonly identified in the liver and adrenal glands, suggesting that these genes may regulate heat stress response through interactions between the liver and adrenal glands. In conclusion, this study would enhance our understanding of the complex underlying mechanisms of acute heat stress, and the identified mRNA isoforms and genes can be used as potential candidates for thermotolerance selection in mammals.The molecular mechanisms underlying heat stress tolerance in animals to high temperatures remain unclear. This study identified the differentially expressed mRNA isoforms which narrowed down the most reliable DEG markers and molecular pathways that underlie the mechanisms of thermoregulation. This experiment was performed on Sprague Dawley rats housed at 22 °C , and three acute heat-stressed groups housed at 42 °C for 30 min (H30), 60 min (H60), and 120 min (H120). Earlier, we demonstrated that acute heat stress increased the rectal temperature of rats, caused abnormal changes in the blood biochemical parameters, as well as induced dramatic changes in the expression levels of genes through epigenetics and post-transcriptional regulation. Transcriptomic analysis using RNA-Sequencing (RNA-Seq) data obtained previously from blood (CT and H120), liver , and adrenal glands  was performed. The differentially expressed mRNA isoforms (DEIs) were identified and annotated by the CLC Genomics Workbench. Biological process and metabolic pathway analyses were performed using Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. A total of 225, 5764, and 4988 DEIs in the blood, liver, and adrenal glands were observed. Furthermore, the number of novel differentially expressed transcript lengths with annotated genes and novel differentially expressed transcript with non-annotated genes were 136 and 8 in blood, 3549 and 120 in the liver, as well as 3078 and 220 in adrenal glands, respectively. About 35 genes were involved in the heat stress response, out of which,  As the global temperature increases, heat stress has become a big challenge for human health and animal survival. An assessment report released by the Intergovernmental Panel on Climate Change (IPCC) predicts that by 2100, the global average surface temperature will rise by 0.3–4.8 °C ,6. AdditTLR4 gene was spliced and 167 bp shorter in the alternative splicing variant ; the response to cold ; the response to oxidative stress .In the liver, the novel transcript lengths of the annotated genes were found to be down-regulated (72.33% of 3549 in three comparisons), and 187 novel transcript lengths of the annotated known genes were shared among the CT vs. H30, CT vs. H60, and CT vs. H120 comparisons . Moreovep ≤ 0.001, FDR ≤ 0.05, and |FC| ≥ 2, and 156 genes were commonly identified in three comparisons, including 63 genes that were up-regulated and 93 genes that were down-regulated when heat stress occurred. We also found that most shared genes were engaged in the response to hormones, the response to heat, and metabolic processes, such as Btg1 (BTG anti-proliferation factor 1), Lrp5 (LDL receptor-related protein 5), Cyp11b3 , Nr4a3 , Dnaja1 (DnaJ heat shock protein family (Hsp40) member A1), Dnaja4 (DnaJ heat shock protein family (Hsp40) member A4), Hspd1 and Hsf1 (heat shock transcription factor 1), etc.In adrenal glands , a totalThe differentially expressed novel transcripts associated with non-annotated genes are shown in Gene_338_1 and Gene_786_1 were down-regulated and presented 100% similarity with Calm2 and Gypa , when H30, H60, and H120 were compared to the CT groups, respectively. The Gene_541, located at the genome position “3:163817141-16381790”, had the greatest degree of change among the H30 (FC = −46.82), H60 (FC = −44.03), and H120 (FC = −43.04) treatment groups. A total of 35 differentially expressed novel transcripts were obtained in the H120 conditions in the adrenal glands, which is 1.46 and 5 times more than those transcripts in the H60 and H30 conditions , including Fn1, Rpl11, Dnajb2, etc., which also have the functions of the positive regulation of T cell migration, the negative regulation of the apoptotic process, as well as the negative regulation of I-kappaB kinase/NF-kappaB (NF-κB) signaling. Heat stress is well documented to have a negative impact on the immune system through humoral immune responses and cell-mediated responses [Based on FDR ≤ 0.05, 77 BP terms were found in the blood for the CT vs. H120 comparison, and most of them were related to the immune response, such as the regulation of adaptive immune response (GO:0002819) and the regulation of T cell-mediated immunity (GO:0002709) . Furtheresponses ,74. In tp ≤ 0.05) in the response to arsenic-containing substances (2) clustered by 14 genes that were specifically identified in the CT vs. H120 comparison  BP terms were detected in the CT vs. H30, CT vs. H60, and CT vs. H120 comparisons , respectbstances C. In addmparison B, and thIgfbp7, Dnaja1, LOC680121 (similar to heat shock protein 8), Bag3, Hsp90aa1, and Hspd1 were shared in the CT vs. H30, CT vs. H60, and CT vs. H120 comparisons (Hsf1 and Trpv2 genes were significantly enriched both in the response to heat- and cold-induced thermogenesis processes at H60 and H120 (p ≤ 0.01) roles in protein folding and the positive regulation of reactive oxygen species metabolic processes (p ≤ 0.01) involved in the regulation of transcription from the RNA polymerase H promoter in response to stress, sterol homeostasis, and the regulation of DNA-template transcription in response to stress  detected in the CT vs. H30, CT vs. H60, and CT vs. H120 comparisons, respectively . Among tparisons A. There parisons B. Furtheand H120 A,B. Furtrocesses C. Genes o stress D.HSP70 (HSP72), HSP90, HSF-1, and TRP channels [HSF-1 was induced by cold shock at 4 °C, and then increased the expression of HSP70 and HSP90 by binding to the heat response element [HSF-1 not only mediated the expression of HSP during the period of heat stress but also after rewarming to 37 °C. The TRP channels are often activated by a variety of stressors . TRPV1-4 and TRPM2 are activated by heat stress, and TRPA1 and TRPM8 are activated by cold stress [HSF-1 and Trpv2 were found to be involved in the process of the response to heat- and cold-induced thermogenesis at H60 and H120, suggesting that both HSF-1 and Trpv2 played important roles in thermoregulation, but further validation experiments are needed to confirm their functions. The Trpv2 gene can be considered as a novel target for a subsequent heat stress study.It is widely known that changes in gene expression may be triggered by thermal stress; in turn, the expression of genes may help cells to defend themselves against thermal stress. Some genes were both activated by heat and cold stress, such as the channels . A previ element . This std stress . In the 10), which clustered 13 genes, including RT1-Ba, Prf1, RT1-CE5, RT1-CE7, RT1-T24-1, RT1-Bb, RT1-Da, RT1-Db1, Actb, Actg1, RT1-CE4, RT1-CE10, and LOC108348139 (Thirty-three metabolic pathways were significantly detected in the blood, which were divided into five categories, including the cellular process, the environment information process, human diseases, metabolism, and organismal systems . A total08348139 . Kanda eIn the liver, 31, 46, and 51 pathways were significantly enriched in the CT vs. H30, CT vs. H60, and CT vs. H120 comparisons, respectively . Eleven Dnaja1, LOC680121, Chordc1, AABR07011951.1, Hsp90aa1, Hspa1b, Cdkn1a, Hmox1, Bag3, and Dnaja4, suggesting that these genes may regulate the heat stress response through crosstalk between the liver and adrenal glands. Combining with the expression profiles from the RNA-Seq and the validation results of the RT-qPCR experiments performed in our previous study [A total of 43, 786, and 1149 DEIDEGs were found in blood, liver, and adrenal gland tissues under heat stress, respectively . Genes eUnderstanding mechanisms of heat stress response and discovering indicators to quantify the degree of heat stress are challenging for researchers ,83. NumeThis research described the transcriptome profiles in the blood, liver, and adrenal glands of Sprague Dawley rats under various heat stress conditions, showing that the expression of heat stress-responsive genes in rats has spatiotemporal differences. In total, 35 genes were involved in the process of thermal stress regulation annotated by DEIs and were also differentially expressed in the liver and adrenal gland tissues, which could be preferentially considered as candidate markers for heat stress in rats. Furthermore, the identification of novel mRNA isoforms related to heat stress response allows for new insights into a better understanding of transcripts functionally playing important roles in heat stress, which may in the future be used to select for thermotolerance. The evident role of the inflammation–metabolism nexus appears to be central in the acute-heat-stressed rats through mRNA isoform identification, and hence this study indirectly emphasizes this intervention nexus for heat stress management. The magnitude of transcriptional changes uncovered in this study points towards the need for pathway-level biological investigation, along with further studies on the essential crosstalk between different biological pathways. Future GWAS studies focusing on the thermotolerance breeding of livestock can benefit from this study in the selection of genes, and a careful approach taking into account the spatiotemporal and post-transcriptional changes can be adopted.


Accumulating evidence has suggested the possibility of FTO and ALKBH5 as therapeutic targets for specific diseases. In this review, we aim to illustrate the structural properties of these two m6A demethylases and the development of their specific inhibitors. Additionally, this review will summarize the biological functions of these two m6A demethylases in various types of cancers and other human diseases.N6-methyladenosine (m6A) is a post-transcriptional RNA modification and one of the most abundant types of RNA chemical modifications. m6A functions as a molecular switch and is involved in a range of biomedical aspects, including cardiovascular diseases, the central nervous system, and cancers. Conceptually, m6A methylation can be dynamically and reversibly modulated by RNA methylation regulatory proteins, resulting in diverse fates of mRNAs. This review focuses on m6A demethylases fat-mass- and obesity-associated protein (FTO) and alkB homolog 5 (ALKBH5), which especially erase m6A modification from target mRNAs. Recent advances have highlighted that FTO and ALKBH5 play an oncogenic role in various cancers, such as acute myeloid leukemias (AML), glioblastoma, and breast cancer. Moreover, studies in vitro and in  Existing epigenetics mainly involve reversible chemical modifications of DNA, histone, and RNA, which can be inherited through cell division without changing the DNA sequence. So far, over 100 kinds of post-transcriptional modifications of RNA have been defined, including N1-methyladenosine (m1A), N6-methyladenosine (m6A), and 2-O-dimethyladenosine (m6Am) , and was2+. They all belong to the AlkB subfamily of the 2OG dioxygenase superfamily . O39, 40, 41, 50 values of 0.84 μM and 1.79 μM, respectively [50 ranging from 1.38 to 16.5 μM. In addition, some FTO inhibitors also showed similar activity with ALKBH5. For example, amiloride analogue 25a (25c-2 (25e-3 (50 values of 1.98 μM, 7.13 μM, and 1.24 μM for target ALKBH5, respectively.The imidazobenzoxazin-5-thione MV1035 (compound ectively . This stogue 25a , rhein aa (25c-2 , and com2 (25e-3  showed iNumerous epigenetic studies in recent decades have revealed the potential of m6A demethylases as therapeutic targets for Human Diseases, including cancers. However, these two demethylases are likely to display different effects on various diseases because they are abundant in different tissues and have a different selection of action sites. Given the above, FTO is intensely related to a range of biomedical processes, including obesity-related diseases, metabolic homeostasis, pro-tumor, self-renewal ability of CSCs, immunotherapy resistance, and chemotherapy resistance. Correspondingly, ALKBH5 shows a clear association with pro-tumor, cancer suppression, self-renewal ability of CSCs, autophagy, chemotherapy resistance, and other diseases. Nevertheless, few studies have reported on the regulation of cellular contexts, such as immunity, DNA damage, autophagy, and apoptosis by m6A demethylases. Thus, future work is certainly required to determine the regulatory genes of each m6A demethylase in various cancers.mice. However, few of the currently developed small-molecule FTO inhibitors are available for clinical applications due to mild bioavailability, low sensitivity, and/or poor selectivity. Therefore, much work remains to be carried out in the future to develop more potent FTO inhibitors and to improve their biological functions, inhibitory effects, and the therapeutic potential for human disease treatment. In addition, designing and synthesizing FTO inhibitors based on existing small molecules, and discovering FTO inhibitors with distinct frameworks or compounds that bind to novel binding sites, should be considered as an important research strategy in the future work. Additionally, the lack of selectivity was a challenge for ALKBH5 inhibitors. One significant hindrance is its flexible motif 2 modification and unique disulfide bond, resulting in the smaller active pocket and smaller inhibitors. Therefore, identifying new and lead ALKBH5 inhibitors is urgently needed.At present, reported FTO/ALKBH5 inhibitors mainly focus on 2OG analogs and substrate-competitive inhibitors. Several potent FTO inhibitors have been demonstrated to suppress the proliferation of cancer cells, such as R-2HG, FB23-2, CS1, CS2, and SsD in leukemia cells ,118,122;In conclusion, this review underlines the recent advances of m6A demethylases in many Human Diseases. However, there are still some issues that should be resolved. Firstly, the underlying mechanisms of m6A demethylases in some cancers are not fully understood. Secondly, some findings have shown that m6A demethylases can be used as therapeutic targets, but specific experiments in clinical trials remain to be conducted. Thirdly, the possibility of m6A demethylase inhibitors for further clinical applications, or in combination with clinical drugs for specific diseases, should be carefully explored.