Search Results (4,755)

Candida albicans is a major cause of human mucosal and invasive fungal infections, but the relationship between its intraspecific genomic diversity and metabolic variation remains poorly understood. Here, we integrated 80 public C. albicans genome assemblies, published fungal genome-scale metabolic models (GEMs), public reaction databases, and orthogroup-linked gene–protein–reaction (GPR) evidence to construct a species-level C. albicans pan-GEM and derived 80 strain-specific GEMs (ssGEMs) through genome projection. The pan-genome comprised 10,308 orthogroups, including 4215 core, 5947 accessory, and 146 singleton orthogroups. The final pan-GEM contained 1986 reactions, 1777 metabolites, and 865 genes. After feasibility rescue, all 80 ssGEMs met the feasibility criterion for predicted growth and passed the closed-uptake energy-generating-cycle test. Among experimentally essential genes with resolvable GPR associations, 23 were consistently predicted as model-essential across all final ssGEMs. As an application of the ssGEM collection, nutrient-boundary simulations showed that increasing D-glucose uptake markedly increased predicted growth across 79 feasible ssGEMs. This framework provides a reusable resource for comparing conserved metabolic functions and genome-projected reaction differences across C. albicans strains. Full article

Background/Objectives: Polycystic ovary syndrome (PCOS) is heterogeneous, and oral hygiene indices may not capture the microbial and inflammatory features relevant to different phenotypes. We evaluated oral hygiene, salivary microbiota, salivary zonulin, and systemic biomarkers in classical PCOS and non-classical PCOS (Rotterdam phenotypes C and D). Methods: This cross-sectional analysis used 110 complete records from the master dataset: 42 women with classical PCOS, 45 with non-classical PCOS, and 23 controls. Oral hygiene was assessed using the Oral Hygiene Index-Simplified (OHI-S). Serum interleukin-1 beta (IL-1β), anti-Müllerian hormone (AMH), fasting glucose, and insulin were assessed, salivary zonulin was measured by ELISA, and salivary Lactobacillus, Prevotella, and Bifidobacterium profiles were evaluated by qPCR. Spearman correlations were tested as an exploratory family of 24 phenotype-specific comparisons using the Benjamini–Hochberg false discovery rate (FDR); a BMI-adjusted sensitivity analysis was also performed for the debris–zonulin association. Results: OHI-S indices and HOMA-IR did not differ among groups. Raw between-group differences for IL-1β and AMH did not remain significant after FDR correction. Prevotella and Bifidobacterium qPCR-derived abundance differed among groups, whereas Lactobacillus did not. In non-classical PCOS, the Debris Index correlated positively with salivary zonulin (ρ = 0.46, raw p = 0.002, FDR q = 0.036; bootstrap 95% CI 0.14–0.73); the BMI-adjusted partial Spearman correlation was similar (ρ = 0.47, p = 0.001). The inverse calculus–zonulin association in classical PCOS and the total OHI-S–zonulin association in non-classical PCOS did not remain significant after FDR correction. Conclusions: OHI-S was not a discriminatory marker of PCOS status in this cohort. One phenotype-specific debris–zonulin association remained after correction, but the cross-sectional design, restricted OHI-S range, and small control group require cautious interpretation. These findings support further hypothesis-driven studies rather than a clinical claim for an established oral–gut axis in PCOS. Full article

Coffee is one of the most widely consumed beverages worldwide and a major dietary source of bioactive compounds, including caffeine, chlorogenic acids (CGAs), trigonelline, non-digestible polysaccharides, and roasting-derived melanoidins. Habitual coffee consumption has been associated with a lower risk of type 2 diabetes (T2D) and more favorable liver-related outcomes, whereas evidence regarding obesity and metabolic dysfunction-associated steatotic liver disease (MASLD) remains heterogeneous. These associations may reflect interactions among coffee constituents, the gut microbiota, and host metabolic phenotypes rather than the effects of individual compounds alone. In this focused narrative review, we integrate evidence on the gastrointestinal fate, microbial transformation, and systemic availability of major coffee constituents, with particular emphasis on short-chain fatty acids, CGA-derived phenolic acids, and microbiota-dependent bile acid metabolism. We examine how these pathways may influence intestinal barrier integrity, mucosal immunity, enteroendocrine signaling, and gut–liver communication then consider their potential relevance to the regulation of energy, glucose, and lipid metabolism. We evaluate the relevance of these pathways to obesity, T2D, and MASLD, as well as major sources of inter-individual variability, including host genetics, baseline microbiota, metabolic phenotypes, background diet, and coffee composition, processing, brewing, filtration, dose, and beverage additives. Finally, we discuss physiologically relevant exposure levels, candidate biomarkers of individual responses, and risk–benefit considerations. Although mechanistic and preclinical evidence is accumulating, human studies directly demonstrating that coffee-induced microbiota changes mediate beneficial metabolic outcomes remain limited. Future randomized and mechanistic studies should integrate chemically characterized coffee exposures with longitudinal microbiome, metabolomic, physiological, and clinically relevant outcome measurements. Full article

The hypothalamus is a central regulator of systemic energy balance and glucose homeostasis, yet how type 2 diabetes (T2D) alters its cellular and regulatory landscape remains incompletely understood. Here, we integrated single-nucleus RNA sequencing (snRNA-seq) with bulk ATAC-seq to characterize hypothalamic remodeling in an HFD/STZ-induced T2D mouse model. High-resolution snRNA-seq revealed pronounced cell type-specific alterations involving several neuronal subpopulations, including Gal.Nts.Npy, Nxph4.Adcyap1, and Lmx1a.Gpr149.Tcf4 neurons, together with substantial changes in oligodendrocyte-lineage cells. Increased Npy expression was accompanied by greater tissue-level chromatin accessibility at the Npy locus, providing convergent evidence for activation of the hypothalamic Npy regulatory axis. Pseudotime analysis further showed increased representation of a transcriptionally distinct Cntnap2-positive oligodendrocyte-lineage state, together with reduced representation of canonical myelination-associated states. Ligand–receptor analysis predicted reduced signaling across selected neuron–oligodendrocyte interactions and altered communication among glial populations. Collectively, these findings reveal coordinated neuronal, oligodendrocyte-lineage, intercellular signaling, and chromatin-accessibility alterations in the diabetic hypothalamus, and identify the Npy regulatory axis and Cntnap2-positive oligodendrocyte-lineage state as candidate features for further mechanistic investigation. Full article

Background/Objectives: Despite the rapid development of personalised nutrition, new tools are still needed to support and improve diet quality and prevent diet-related diseases in individuals who do not seek or require personalised dietary advice. The study analysed changes in diet quality, body composition and cardiometabolic markers in two groups of the Dietary Diary app (DDApp) users: (i) after 10 weeks in invited app users, (ii) after one year in voluntary app users who, after 10 weeks, were not encouraged to continue using the app. Methods: The app underwent testing with 62 adult participants (aged 18–69 years). App users’ body composition, systolic (SBP) and diastolic (DBP) blood pressure, total cholesterol (TC) and fasting blood glucose in capillary blood (FBG) were measured before (62 users), after 10 weeks (62) and after a year (30). Dietary data were collected, and pro-Healthy Diet Index (pHDI) and non-Healthy Diet Index (nHDI) were calculated. Results: After 10 weeks, invited app users recorded decreased nHDI (mean ± standard deviation: from 15.5 ± 6.2 to 12.1 ± 4.9% points; p < 0.001), body weight (81.5 ± 18.3 to 79.7 ± 18.0 kg; p < 0.001), body mass index (28.6 ± 6.1 to 28.0 ± 6.1 kg/m²; p < 0.001), waist circumference (91.2 ± 14.8 to 89.7 ± 14.4 cm; p = 0.019), fat content (36.6 ± 7.9 to 35.1 ± 8.5% bw.; p < 0.001), FBG (108.3 ± 30.1 to 93.5 ± 17.3 mg/dL; p < 0.001), TC (199.1 ± 36.6 to 169.3 ± 25.5 mg/dL; p < 0.001), SBP (131.3 ± 16.5 to 123.5 ± 15.6 mmHg; p < 0.001), DBP (84.8 ± 9.8 to 80.1 ± 9.3 mmHg; p < 0.001), along with increased fat-free mass/fat mass ratio (1.9 ± 0.9 to 2.1 ± 0.9; p < 0.001) and muscle mass/fat mass ratio (0.9 ± 0.5 to 1.0 ± 0.5; p < 0.001). After a year, voluntary app users had decreased nHDI (from 16.7 ± 6.7 to 13.5 ± 6.1% points; p = 0.017), visceral fat tissue (1.9 ± 1.8 to 1.8 ± 1.7 kg; p = 0.045) and SBP (129.9 ± 15.9 to 125.0 ± 13.5 mmHg; p = 0.015). Conclusions: DDApp may support dietary behaviour change through self-monitoring, with favourable short-term reductions in nHDI and improvements in body composition and selected cardiometabolic markers. Some benefits persisted after one year. Full article

Primitive Myeloid Cells (PMCs) arise from the anterior ventral blood islands during Xenopus laevis embryogenesis and migrate to colonize embryonic tissues, contributing to wound healing, innate immunity, and development. Because cell movement and growth are metabolically demanding, we investigated whether metabolic pathways regulate PMC behaviour integrating public single-cell RNA-seq data with whole-mount in situ hybridization (WMISH) to profile metabolic gene expression in PMCs, followed by functional assays using pathway-specific inhibitors: 2-deoxy-D-glucose 2-DG (global glucose metabolism), YZ-9 (glycolysis), 6-AN (pentose phosphate pathway, PPP), and ST045849 (hexosamine biosynthetic pathway, HBP). The analyses showed that PMCs express high levels of glycolytic and ancillary metabolic enzymes. Global inhibition impaired PMC colonization and wound recruitment and modestly reduced the size of the specification domain, confirming a requirement for glycolytic flux. PPP and downstream glycolysis inhibition produced only modest, non-significant shifts in recruitment, whereas HBP inhibition abolished wound recruitment and reduced PMC numbers. These findings identify glucose metabolism as a regulator of PMC colonization and immune function, revealing how metabolic reprogramming governs early immune cell behaviour independent of a functional vasculature. Full article

Background: Practical localized resistance tasks involving functionally divergent muscle groups may produce different systemic biochemical responses, although neither task isolates a single muscle. Methods: In this exploratory, nonrandomized, order-balanced within-participant crossover study, 102 young adults completed bilateral seated plantar flexion and unilateral elbow flexion, with 51 participants assigned by alternation to each condition sequence. The analysis-ready biochemical dataset contained complete observations for 102 participants. Blood was collected before and 2–3 min after each condition. We analyzed plasma glucose and lactate and serum total cholesterol, HDL-C, LDL-C, triglycerides, creatine kinase, lactate dehydrogenase, GGT, and cortisol. Linear mixed-effects models included exercise condition, sampling time, study period, assigned sequence, and their respective interactions with sampling time as fixed effects. Participant and participant-specific study visit were included as random intercepts where supported by the model. The exercise condition × time interaction remained the primary contrast, and Holm correction was applied across the ten predefined biochemical outcomes. Linear mixed-effects models included exercise condition, sampling time, study period, assigned sequence, and their respective interactions with sampling time as fixed effects. Participant and participant-specific study visit were included as random intercepts where supported by the model. The exercise condition × time interaction remained the primary contrast, and Holm correction was applied across the ten predefined biochemical outcomes. Results: After adjustment for period and sequence and Holm correction across the ten primary interaction tests, cortisol remained the only statistically significant exercise condition × time interaction (149.219 nmol·L⁻¹, 95% CI 62.338 to 236.099; unadjusted p = 0.000821; Holm-adjusted p = 0.008). Glucose showed a negative interaction estimate (−7.013 mg·dL⁻¹, 95% CI −12.238 to −1.788; unadjusted p = 0.009), but did not remain significant after Holm correction (adjusted p = 0.078). No period × time or sequence × time effect remained statistically significant after correction. Sensitivity analyses were not fully concordant with the primary model, supporting cautious interpretation of the cortisol finding. Conclusions: These findings describe immediate responses to two non-equivalent multi-muscle tasks and cannot be attributed to selective soleus or biceps-brachii activation, fiber-type composition, or muscle-specific substrate use. Full article

Peripheral arterial disease (PAD) is a major cardiovascular complication of hemodialysis (HD), during which inflammation and oxidative stress contribute to vascular injury. The role of spermine oxidase (SMOX), a polyamine-metabolizing enzyme generating hydrogen peroxide, in PAD is unknown in HD. This cross-sectional study included 142 patients on maintenance HD, including 115 with a normal ankle–brachial index (ABI) (≥0.9) and 27 with a low ABI (<0.9). Patients with a low ABI were older and had lower creatinine and albumin levels, fractional clearance index for urea, and urea reduction ratio but higher levels of glucose, C-reactive protein (CRP), and SMOX; diabetes mellitus was also more prevalent (p < 0.05 for all). In Firth-penalized multivariable logistic regression, SMOX (adjusted odds ratio [aOR] per 1 ng/mL increase, 1.063; 95% confidence interval [CI], 1.018–1.115; p = 0.001) and CRP (aOR per 0.1 mg/dL increase, 1.181; 95% CI, 1.079–1.321; p < 0.001) were independently associated with ABI-defined PAD. Conventional and penalized sensitivity analyses yielded similar estimates. The areas under the curve for SMOX and CRP were 0.790 and 0.804, respectively. SMOX was inversely correlated with bilateral ABI and positively correlated with CRP. SMOX may represent a candidate biomarker associated with ABI-defined PAD in patients undergoing HD, although further validation in larger prospective cohorts is required. Full article

Reduced expression of MAPK organiser 1 (MORG1) exerts renoprotective effects in MORG1^+/− mice in models of type 1 and type 2 diabetes. To investigate the regulation of MORG1 expression, we generated endogenous hMORG1-luciferase-tagged HEK293 cells carrying a firefly luciferase reporter inserted downstream of the human MORG1 gene using the CRISPR/Cas9 method. This study analysed the time-dependent effects of 20 mM D-glucose on MORG1-luciferase activity and examined whether 20 µM metformin influences MORG1 expression. HEK293 cells were genomically modified using CRISPR/Cas9 to insert the firefly luciferase gene downstream of MORG1. Stable clones were selected with puromycin. The influence of D-glucose concentration on MORG1 expression was analysed in a dose- and time-dependent manner. Luciferase assays assessed MORG1 transcriptional activity, while Western blotting determined protein expression. qRT-PCR was used to assess MORG1 mRNA levels. The pharmacological effect of metformin was evaluated in parental and tagged HEK293 cells and TKPTS cells, simultaneously exposed to 5.5 mM or 20 mM D-glucose in the presence or absence of 20 µM metformin. High glucose significantly increased luciferase activity and MORG1 protein expression compared with 5.5 mM glucose controls. Time course experiments demonstrated significant induction of MORG1 transcription, confirmed by corresponding increases in protein levels. Metformin treatment significantly reduced glucose-induced luciferase activity, MORG1 mRNA levels, and MORG1 protein expression at 24 h. Analyses of TKPTS cells show that 20 mM D-glucose elevated Morg1 mRNA levels, while this induction was attenuated in the presence of 20 µM metformin. High glucose induces time-dependent upregulation of MORG1 transcription and protein expression in MORG1-luciferase-tagged HEK293 cells. Metformin application attenuates, but does not fully prevent, glucose-induced MORG1 mRNA and proteinexpression. Full article

Major burn injury induces stress-related hyperglycemia, but absolute glucose concentrations do not account for chronic glycemic status. The glycemic gap, defined as the difference between measured glucose and hemoglobin A1c (HbA1c)-derived estimated average glucose, may reflect acute glycemic deviation from chronic glycemia. We investigated the association between preoperative glycemic gap and 30-day postoperative bloodstream infection (BSI) in adult burn intensive care unit patients without diabetes. In this retrospective cohort study, 352 patients undergoing burn surgery between 2014 and 2024 were included, of whom 217 (61.6%) developed postoperative BSI. Each 10-mg/dL increase in preoperative glycemic gap was independently associated with higher odds of BSI (odds ratio, 1.104; 95% confidence interval, 1.036–1.184; p = 0.003) after adjustment for clinical factors, including burn severity. This association remained consistent in a sensitivity analysis using the revised Baux score. Adding either preoperative glucose or glycemic gap to the clinical model improved model fit, whereas improvements in discrimination were modest, with no significant difference between the glucose and glycemic gap models. A greater preoperative glycemic gap was independently associated with subsequent BSI in patients with major burns without diabetes and may provide complementary information regarding acute glycemic disturbance. Full article

Background/Objectives: Subtle cognitive alterations have been described in children with type 1 diabetes (T1D), but the association between cognition and real-time glycemia during cognitive testing remains unclear. This study aimed to investigate whether blood glucose level (BGL) measured immediately before the MOXO Continuous Performance Test (MOXO-CPT) is associated with cognitive performance and modified by clinical factors. Methods: A total of 117 children with T1D (aged 6–18 years) completed the MOXO-CPT assessing sustained attention, timeliness, impulsivity, and hyperactivity. Participants were stratified by BGL measured immediately before testing (<200 mg/dL [<11.1 mmol/L] vs. ≥200 mg/dL [≥11 mmol/L]). Group comparisons were performed using the Mann–Whitney U test. Multivariable linear regression models examined associations between BGL and cognitive outcomes, adjusting for HbA1c, age, sex, treatment modality and diabetes duration. Interaction analyses were also conducted. Results: In unadjusted group comparisons, participants with BGL ≥ 200 mg/dL demonstrated nominally higher sustained attention (p = 0.036) and timeliness scores (p = 0.040) than those with BGL < 200 mg/dL. However, these differences did not remain statistically significant after Benjamini–Hochberg correction and were not confirmed after adjustment for relevant covariates. No differences were observed in impulsivity or hyperactivity. In multivariable models, higher BGLs were independently associated with higher impulsivity scores (B = 0.013, p = 0.007), with higher scores indicating better impulse-control performance. A significant BGL–sex interaction was observed for timeliness. Sex-stratified analyses showed that higher BGLs were associated with better attention and timeliness in girls, whereas the association with impulsivity observed in boys was not confirmed using robust analysis. No significant interactions between BGL and HbA1c, age, treatment modality, or diabetes duration were observed after robust analysis. Conclusions: Real-time glycemia during cognitive testing was associated with subtle, domain-specific differences in cognitive performance in children with T1D. These findings do not establish causality or indicate that higher glucose levels improve cognitive performance. Concurrent glucose levels may be worth considering when interpreting cognitive assessments, although this should be confirmed in prospective studies. Full article

Background/Objectives: Impaired masticatory function may compromise nutritional status in older adults wearing removable dentures, but its associations with nutritional risk indices remain unclear. This study aimed to examine associations between masticatory performance and risk defined by the Geriatric Nutritional Risk Index (GNRI), Prognostic Nutritional Index (PNI), and Controlling Nutritional Status (CONUT) score. Methods: This cross-sectional study included 179 outpatients aged ≥60 years wearing removable dentures. Masticatory performance was quantified by glucose concentration after chewing a gummy jelly. GNRI-defined risk was the prespecified primary outcome; PNI- and CONUT-defined risk were secondary. Logistic regression used Model 1 adjusted for age, sex, and comorbidity and an exploratory Model 2 with additional oral, dietary, and socioeconomic covariates. Results: Mean age was 74.1 ± 6.9 years, and 60.9% were female. Nutritional risk was identified in 21.8%, 48.6%, and 31.3% using the GNRI, PNI, and CONUT, respectively. Higher masticatory performance, indicated by higher glucose concentration, was associated with lower odds of GNRI-defined nutritional risk in both the primary parsimonious model (OR per 10 mg/dL increase, 0.89; 95% CI, 0.82–0.95; p = 0.001) and the exploratory additionally adjusted model (OR, 0.89; 95% CI, 0.82–0.96; p = 0.002). Associations with PNI- and CONUT-defined risk were not consistent across models. Conclusions: Lower masticatory performance was consistently associated with higher GNRI-defined risk, whereas comparable associations were not consistently observed for PNI- or CONUT-defined risk. Objective assessment of masticatory performance may provide clinically relevant information regarding GNRI-defined nutritional risk in older removable denture wearers. Full article

Background/Objectives: Postprandial glycemic responses vary markedly among individuals, but it remains unclear whether the glycemic effect of incremental carbohydrate exposure is systematically modified by insulin resistance. We examined whether insulin resistance alters the carbohydrate–postprandial glucose dose-response across distinct meal contexts. Methods: We performed a secondary repeated-measures analysis of the publicly available CGMacros cohort. Forty-five adults spanning normoglycemia, prediabetes, and type 2 diabetes wore blinded FreeStyle Libre Pro and Dexcom G6 Pro continuous glucose monitors while recording meals for approximately 10 days. The primary outcome was 2-h incremental area under the glucose curve (iAUC). Mixed-effects models tested carbohydrate × HOMA-IR interactions, adjusted for protein, fat, fiber, premeal glucose, age, sex, and BMI. Breakfast was the primary structured context, lunch provided within-cohort contextual confirmation, dinner was a free-living contrast, and Dexcom analyses assessed cross-device robustness. Results: The primary Libre analysis included 423 breakfasts and 414 lunches from 44 participants. Higher HOMA-IR significantly amplified the iAUC associated with each 10-g increment in carbohydrate during breakfast (interaction β = 178.5 mg·min/dL per 1-SD higher HOMA-IR; 95% CI 79.5–277.5; p < 0.001) and lunch (β = 109.8; 95% CI 34.2–185.3; p = 0.004). The direction was supported with Dexcom during breakfast (β = 165.3; 95% CI 45.0–285.6; p = 0.007) and lunch (β = 87.9; 95% CI 2.6–173.1; p = 0.043) and remained robust in GEE, participant fixed-effects, random-slope, activity-adjusted, and other sensitivity analyses. No positive interaction was observed during dinner. Conclusions: In this observational secondary analysis, insulin resistance modified the glycemic impact of carbohydrate across two structured meal contexts, with supportive cross-device evidence. These findings are hypothesis-generating and do not establish HOMA-IR-based dietary prescriptions; prospective external validation is required. Full article

This study evaluated the effects of dietary SYNSEA Premium postbiotics on growth, immunity, disease resistance, intestinal microbiota, and host metabolism in Asian seabass (Lates calcarifer). Fish were fed a control diet or diets supplemented with heat-killed Lactiplantibacillus plantarum LP28, L. plantarum LP1008, and Bacillus subtilis at 10⁸ (LSP) or 10⁹ (HSP) cells kg⁻¹ diet for 56 days. Postbiotic supplementation did not significantly affect growth performance, feed efficiency, production, condition factor, or dorsal muscle composition, but significantly improved survival. Fish receiving postbiotics also exhibited higher survival following Vibrio alginolyticus and iridovirus challenges. These protective effects were accompanied by enhanced superoxide dismutase, phagocytic, and lysozyme activities and modulation of immune-related genes, including tgf-β1, tnf, ifn-γ1, c3, and mx. Exploratory microbiome and metabolome analyses, which were restricted to the control and LSP groups, identified differences in the relative abundance of specific intestinal microbial taxa and associations between microbial composition and host metabolic profiles. The LSP group showed lower relative abundances of potential pathogens such as Salmonella enterica, Lactococcus garvieae, and Staphylococcus warneri, although the overall microbial community structure did not differ significantly between groups. Metabolomic analysis of the LSP group further showed changes in D-glucose, pentose phosphate pathway intermediates, reduced glutathione, CoA, and 2-methylacetoacetyl-CoA relative to the control. Collectively, SYNSEA Premium improved survival, immune responses, and resistance to bacterial and viral infections without significantly affecting growth performance, while exploratory omics analysis of the LSP treatment identified associated microbial and metabolic changes. Full article

Seed dormancy is an important adaptive mechanism in plants. However, some seeds, such as those of Xanthoceras sorbifolium (a valuable economic and medicinal species), exhibit strong dormancy, resulting in a very low natural germination rate. This work investigated the mechanism by which low temperatures (LT: −20 °C storage for 60 days) release seed dormancy and promote germination. This was achieved by examining seed germination conditions, applying scanning electron microscopy (SEM), and measuring physiological indicators of seeds and the hormone levels. Targeted metabolomic analysis of sugar and fatty acid metabolism was also performed. The results were as follows: (1) A high germination rate of 47.3% was observed under LT condition, compared to 32.7% at room temperature (RT: 25 °C). Of four germination methods, direct GMS (germination in moist sand) was the most effective. (2) The two stages of VI (after storage) and VII (on the 7th day of germination) were key stages to breaking the seed dormancy and triggering germination. At both stages, the high integrity of the seed shells and kernels—particularly, the kernels—was observed using SEM. The activities of SOD, CAT, and POD, as well as the levels of IAA and IPA, and the ratios of IAA/ABA and (IAA + GA3 + ZR + IPA)/ABA (tHor/ABA) were found to be higher, especially in stage VII, where tHor/ABA increased by 62.7%, while ABA decreased by 23.2% in the LT treatment compared to the RT treatment. An optimal germination condition (GMS) created a suitable microenvironment, and this could have maintained highly active antioxidant enzymes and kept H₂O₂ (one of reactive oxygen species, or ROS) within signal transduction levels, and cross-talk to hormones. These changes (enzymes, hormones, ROS, and microenvironment) ensured the seeds reaching an optimal state for dormancy release in the VI stage, and facilitated the seed germination in the VII stage. (3) LT treatment promoted the degradation of starch and fats in the seeds. Significant accumulations of eight soluble sugars, such as glucose, D-fructose, and trehalose, as well as three fatty acids, such as Cis-11,14,17-eicosatrienoic acid (C20-3n3) and γ-linolenic acid (C18-3n6), were observed, while two soluble sugars and one fatty acid decreased under LT conditions. In conclusion, low temperature, as an external signal, together with ROS and GA-ABA-IAA etc. the internal signals co-regulated the seed dormancy release and germination. The moist-sand microenvironment was also a key factor in awakening the embryos of X. sorbifolium seeds. Full article