Shifts in the microbial community and metabolome in rumen ecological niches during antler growth.

Antlers are hallmark organ of deer, exhibiting a relatively high growth rate among mammals, and requiring large amounts of nutrients to meet its development. The rumen microbiota plays key roles in nutrient metabolism. However, changes in the microbiota and metabolome in the rumen during antler growth are largely unknown. We investigated rumen microbiota (liquid, solid, ventral epithelium, and dorsal epithelium) and metabolic profiles of sika deer at the early (EG), metaphase (MG) and fast growth (FG) stages. Our data showed greater concentrations of acetate and propionate in the rumens of sika deer from the MG and FG groups than in those of the EG group. However, microbial diversity decreased during antler growth, and was negatively correlated with short-chain fatty acid (SCFA) levels. Prevotella, Ruminococcus, Schaedlerella and Stenotrophomonas were the dominant bacteria in the liquid, solid, ventral epithelium, and dorsal epithelium fractions. The proportions of Stomatobaculum, Succiniclasticum, Comamonas and Anaerotruncus increased significantly in the liquid or dorsal epithelium fractions. Untargeted metabolomics analysis revealed that the metabolites also changed significantly, revealing 237 significantly different metabolites, among which the concentrations of γ-aminobutyrate and creatine increased during antler growth. Arginine and proline metabolism and alanine, aspartate and glutamate metabolism were enhanced. The co-occurrence network results showed that the associations between the rumen microbiota and metabolites different among the three groups. Our results revealed that the different rumen ecological niches were characterized by distinct microbiota compositions, and the production of SCFAs and the metabolism of specific amino acids were significantly changed during antler growth.

Involvement of Fgf2-mediated tau protein phosphorylation in cognitive deficits induced by sevoflurane in aged rats.

OBJECTIVE: Anesthetics have been linked to cognitive alterations, particularly in the elderly. The current research delineates how Fibroblast Growth Factor 2 (Fgf2) modulates tau protein phosphorylation, contributing to cognitive impairments in aged rats upon sevoflurane administration. METHODS: Rats aged 3, 12, and 18 months were subjected to a 2.5% sevoflurane exposure to form a neurotoxicity model. Cognitive performance was gauged, and the GEO database was employed to identify differentially expressed genes (DEGs) in the 18-month-old cohort post sevoflurane exposure. Bioinformatics tools, inclusive of STRING and GeneCards, facilitated detailed analysis. Experimental validations, both in vivo and in vitro, examined Fgf2's effect on tau phosphorylation. RESULTS: Sevoflurane notably altered cognitive behavior in older rats. Out of 128 DEGs discerned, Fgf2 stood out as instrumental in regulating tau protein phosphorylation. Sevoflurane exposure spiked Fgf2 expression in cortical neurons, intensifying tau phosphorylation via the PI3K/AKT/Gsk3b trajectory. Diminishing Fgf2 expression correspondingly curtailed tau phosphorylation, neurofibrillary tangles, and enhanced cognitive capacities in aged rats. CONCLUSION: Sevoflurane elicits a surge in Fgf2 expression in aging rats, directing tau protein phosphorylation through the PI3K/AKT/Gsk3b route, instigating cognitive aberrations.

Microbiota and Metabolite Profiles in the Feces of Juvenile Sika Deer (Cervus nippon) from Birth to Weaning.

The gut microbiota establishment in young ruminants has a profound impact on their adult production performance. However, the critical phase for the succession of the gut microbial composition and metabolic profiles of juvenile sika deer still needs to be further investigated. Here, we analyzed the fecal microbiota and metabolites of juvenile sika deer during the birth (D1), transition (D42), and rumination (D70) periods based on 16S rRNA sequencing and gas chromatography-time-of-flight mass spectrometry (GC-TOF-MS). The results showed that the fecal bacteria and metabolites composition were significantly different in D1 compared to D42 and D70, and the number of OTUs and the Shannon index were significantly higher in D70 than in D1 (p < 0.05). The relative abundances of Lactobacillus, Lactococcus, and Lachnoclostridium showed a significant increase in D1 compared to D42 and D70, whereas the relative abundances of Ruminococcaceae UCG-005, Ruminococcaceae UCG-010, Ruminococcaceae UCG-014, Christensenellaceae R-7, and Eubacterium coprostanoligenes group were significantly decreased in D1 compared to D42 and D70 (p < 0.05). The amounts of serine, phenylalanine, aspartic acid, ornithine, citrulline, creatine, isoleucine, galactose, and ribose in the feces were significantly higher in D1 compared to D42 and D70. In contrast, the concentrations of cortexolone, resveratrol, piceatannol, fumaric acid, alpha-ketoglutarate, glycerol, uracil-5-carboxylic acid, and maleic acid were significantly decreased in D1. The enrichment analysis showed that amino acid metabolism and carbohydrate metabolism were significantly changed in D1 compared to D42 and D70. The glycine, serine and threonine metabolism; alanine, aspartate and glutamate metabolism; arginine biosynthesis; glyoxylate and dicarboxylate metabolism; citrate cycle; and pyruvate metabolism were significantly enriched across the three periods (p < 0.05). In conclusion, our results suggested that the birth-transition period is a critical phase for the gut bacterial community and metabolic function shift in juvenile sika deer.

Impact of deoxynivalenol on rumen function, production, and health of dairy cows: Insights from metabolomics and microbiota analysis.

Deoxynivalenol contamination in feed and food, pervasive from growth, storage, and processing, poses a significant risk to dairy cows, particularly when exposed to a high-starch diet; however, whether a high-starch diet exacerbates these negative effects remains unclear. Therefore, we investigated the combined impact of deoxynivalenol and dietary starch on the production performance, rumen function, and health of dairy cows using metabolomics and 16 S rRNA sequencing. Our findings suggested that both high- and low-starch diets contaminated with deoxynivalenol significantly reduced the concentration of propionate, isobutyrate, valerate, total volatile fatty acids (TVFA), and microbial crude protein (MCP) concentrations, accompanied by a noteworthy increase in NH3-N concentration in vitro and in vivo (P < 0.05). Deoxynivalenol altered the abundance of microbial communities in vivo, notably affecting Oscillospiraceae, Lachnospiraceae, Desulfovibrionaceae, and Selenomonadaceae. Additionally, it significantly downregulated lecithin, arachidonic acid, valine, leucine, isoleucine, arginine, and proline metabolism (P < 0.05). Furthermore, deoxynivalenol triggered oxidative stress, inflammation, and dysregulation in immune system linkage, ultimately compromising the overall health of dairy cows. Collectively, both high- and low-starch diets contaminated with deoxynivalenol could have detrimental effects on rumen function, posing a potential threat to production performance and the overall health of cows. Notably, the negative effects of deoxynivalenol are more pronounced with a high-starch diet than a low-starch diet.

The physiological dissimilarities of Holstein dairy cows with different milk yields.

BACKGROUND: Even if breed, parity, dietary and environmental management are same, dairy cows still have notable differences in milk yield that may be underpinned by physiologic differences. OBJECTIVES: This study aimed to investigate the physiological dissimilarities of dairy cows with different milk yields. METHODS: Thirty cows were sorted into high milk-yielding cows (group H: 58.93±2.31 kg/day), moderate milk-yielding cows (group M: 44.99±0.54 kg/day), and low milk-yielding cows (group L: 24.99±6.83 kg/day) according to milk yield. Blood was collected and serum parameters were assessed. Rumen fluid was collected for the evaluation of rumen fermentation parameters (RFPs) and bacterial community composition (BCC). RESULTS: Serum prolactin, growth hormone, glutathione peroxidase, immunoglobulin A and non-esterified fatty acid had a significantly positive correlation with milk yield (p < 0.05), whereas serum glucagon and total antioxidant capacity had a significantly negative correlation with milk yield (p < 0.05). The concentration of valeric acid and the ratio of acetic acid to propionic acid in the rumen fluid in group H was significantly lower than that in group L (p < 0.05). The concentration of acetic acid and butyric acid in group H was significantly lower than that in groups M and L (p < 0.05). The relative abundances of Ruminococcaceae_NK4A214_group, Prevotella_1, Rikenellaceae_RC9_gut_group, Christensenellaceae_R-7_group, Muribaculaceae, and Ruminococcus_2 were negatively correlated with milk yield, whereas the relative abundance of Succinivibrionaceae_UCG-001, Lachnospiraceae_NK3A20_group, Shuttleworthia and Dialister were positively correlated with milk yield (p < 0.05). CONCLUSIONS: This study indicates that dairy cows with different milk yields have clear divergence in serum indicators, RFPs, BCC and rumen microbial metabolism.

Integrated multi-omics of the gastrointestinal microbiome and ruminant host reveals metabolic adaptation underlying early life development.

BACKGROUND: The gastrointestinal tract (GIT) microbiome of ruminants and its metabolic repercussions vastly influence host metabolism and growth. However, a complete understanding of the bidirectional interactions that occur across the host-microbiome axis remains elusive, particularly during the critical development stages at early life. Here, we present an integrative multi-omics approach that simultaneously resolved the taxonomic and functional attributes of microbiota from five GIT regions as well as the metabolic features of the liver, muscle, urine, and serum in sika deer (Cervus nippon) across three key early life stages. RESULTS: Within the host, analysis of metabolites over time in serum, urine, and muscle (longissimus lumborum) showed that changes in the fatty acid profile were concurrent with gains in body weight. Additional host transcriptomic and metabolomic analysis revealed that fatty acid ß-oxidation and metabolism of tryptophan and branched chain amino acids play important roles in regulating hepatic metabolism. Across the varying regions of the GIT, we demonstrated that a complex and variable community of bacteria, viruses, and archaea colonized the GIT soon after birth, whereas microbial succession was driven by the cooperative networks of hub populations. Furthermore, GIT volatile fatty acid concentrations were marked by increased microbial metabolic pathway abundances linked to mannose (rumen) and amino acids (colon) metabolism. Significant functional shifts were also revealed across varying GIT tissues, which were dominated by host fatty acid metabolism associated with reactive oxygen species in the rumen epithelium, and the intensive immune response in both small and large intestine. Finally, we reveal a possible contributing role of necroptosis and apoptosis in enhancing ileum and colon epithelium development, respectively. CONCLUSIONS: Our findings provide a comprehensive view for the involved mechanisms in the context of GIT microbiome and ruminant metabolic growth at early life. Video Abstract.

Integrated Transcriptome and Microbiota Reveal the Regulatory Effect of 25-Hydroxyvitamin D Supplementation in Antler Growth of Sika Deer.

The level of plasma 25-hydroxyvitamin D (25(OH)D) is associated with the growth of the antler, a fast-growing bone organ of Cervidae. However, the benefits of 25(OH)D supplementation on antler growth and the underlying mechanisms remain unclear. Here, the antler growth profile and transcriptome, plasma parameters, rumen bacteria, and metabolites (volatile fatty acids and amino acids) were determined in sika deer in a 25(OH)D supplementation group (25(OH)D, n = 8) and a control group (Ctrl, n = 8). 25(OH)D supplementation significantly increased the antler weight and growth rate. The levels of IGF-1,25(OH)D and 1,25-dihydroxyvitamin D were significantly higher in the 25(OH)D group than in the Ctrl group, while the levels of LDL-C were lower. The levels of valerate and branched-chain amino acids in the rumen fluid were significantly different between the 25(OH)D and Ctrl groups. The bacterial diversity indices were not significantly different between the two groups. However, the relative abundances of the butyrate-producing bacteria (families Lachnospiraceae and Succinivibrionaceae) and the pyruvate metabolism pathway were higher in the 25(OH)D group. The transcriptomic profile of the antler was significantly different between the 25(OH)D and Ctrl groups, with 356 up- and 668 down-regulated differentially expressed genes (DEGs) in the 25(OH)D group. The up-regulated DEGs were enriched in the proteinaceous extracellular matrix and collagen, while the down-regulated DEGs were enriched in the immune system and lipid metabolism pathways. Overall, these results provide novel insights into the effects of 25(OH)D supplementation on the host metabolism, rumen microbiota, and antler transcriptome of sika deer.

FOXK2 promotes the proliferation of papillary thyroid cancer cell by down-regulating autophagy.

Papillary thyroid cancer (PTC) is the most common endocrine system tumor. FOXK2 is involved in the development of different types of cancers, however, its function has not been investigated in papillary thyroid cancer. In the present study, we demonstrated that FOXK2 expression was up-regulated in papillary thyroid carcinoma tissues compared with matched normal tissues. Importantly, we found that FOXK2 expression was significantly associated with the tumor size, T stage, and TNM stage. Furthermore, stable knockdown of FOXK2 markedly inhibited PTC cell proliferation, significantly increased the ratio of LC3-II/LC3-I, and reduced p62 expression, whereas overexpression of FOXK2 showed opposite effects. In FOXK2 knockdown cell lines, mCherry-GFP-LC3 immunofluorescence demonstrated increased punctate aggregates of mCherry-GFP-LC3, and transmission electron microscopy revealed increased numbers of autophagosomes. Autophagy-related protein ULK1, VPS34, and FOXO3 were markedly up-regulated by FOXK2 knockdown and down-regulated by FOXK2 overexpression. Finally, autophagy inhibitor 3-MA attenuated autophagy activation and rescued the inhibition of cell proliferation caused by FOXK2 knockdown, suggesting that FOXK2 silencing inhibits cell proliferation through up-regulating autophagy. These findings revealed an important role of FOXK2 in PTC progression and suggested that FOXK2 might be a potential new target for the diagnosis and treatment of PTC.

Erratum: FOXK2 promotes the proliferation of papillary thyroid cancer cell by down-regulating autophagy: Erratum.

[This corrects the article DOI: 10.7150/jca.60730.].

Rational Design of ß-NiOOH Nanosheet-Sheathed CNTs as a Highly Efficient Electrocatalyst for Practical Li-S Batteries.

The shuttle effects of polysulfide intermediates (LiPSs) and sluggish kinetics during sulfur reduction reaction (SRR) process severely exacerbate the electrochemical performances of Li-S batteries. Herein, a unique nanocatalyst comprising ß-NiOOH nanosheets uniformly implanted on the surface of carbon nanotubes (CNT@NiOOH) was designed and synthesized for sulfur cathodes. The ß-NiOOH nanosheets have great capability of adsorbing LiPSs as well as superior catalytic activity for accelerating LiPS conversion, providing a more efficient method to restrain shuttle effects and improve the kinetics of SRR. Moreover, the nanometer-scale epitaxial growth and uniform distribution of ß-NiOOH on CNTs provide a multidimensional catalytic skeleton with sufficient accessible active surfaces, unimpeded LiPS diffusion pathways, and resultant high utilization of active sites. Simultaneously, stable electron transportation pathways are also obtained by being synthesized on CNTs to avoid the faultiness of poor electron conductivity of ß-NiOOH. These conspicuous advantages contribute to fully exert the catalytic and LiPS anchoring potential of CNT@NiOOH, bringing about the ultralong cycle performance and excellent capacity reversibility at a high discharge rate. Reticular CNT@NiOOH frameworks are assembled with the sulfur composite materials (SCMs) by a self-assembly method, and a super-high capacity of 813.3 mA h g-1 after 400 cycles at 0.5 C with a small capacity degradation of 0.07% per cycle is achieved. Furthermore, the 3 A h pouch-type cell with the SCM/CNT@NiOOH cathode attains a super-high energy density of about 320 W h kg-1 and shows a superior capacity retention as high as 75.9% after 50 cycles at 0.2 C. This work provides a promising method to accelerate the SRR process and restrain the shuttle effects for practical long-life and high-capacity Li-sulfur batteries.

Effects of Different Combinations of Sugar and Starch Concentrations on Ruminal Fermentation and Bacterial-Community Composition in vitro.

High levels of starch is known to have positive effects on both energy supply and milk yield but increases the risk of rumen acidosis. The use of sugar as a non-structural carbohydrate could circumvent this risk while maintaining the benefits, but its effects and that of the simultaneous use of both sugar and starch are not as well-understood. This study aimed to evaluate the effects of different combinations of sugar and starch concentrations on ruminal fermentation and bacterial community composition in vitro in a 4 ×4 factorial experiment. Sixteen dietary treatments were formulated with 4 levels of sugar (6, 8, 10, and 12% of dietary dry matter), and 4 levels of starch (21, 23, 25, and 27% of dietary dry matter). Samples were taken at 0.5, 1, 3, 6, 12, and 24 h after cultivation to determine the disappearance rate of dry matter, rumen fermentation parameters and bacterial community composition. Butyric acid, gas production, and Treponema abundance were significantly influenced by the sugar level. The pH, acetic acid, and propionic acid levels were significantly influenced by starch levels. However, the interactive effect of sugar and starch was only observed on the rate of dry matter disappearance. Furthermore, different combinations of starch and sugar had different effects on volatile fatty acid production rate, gas production rate, and dry matter disappearance rate. The production rate of rumen fermentation parameters in the high sugar group was higher. Additionally, increasing the sugar content in the diet did not change the main phylum composition in the rumen, but significantly increased the relative abundance of Bacteroidetes and Firmicutes phyla, while the relative abundance of Proteobacteria was reduced. At the genus level, the high glucose group showed significantly higher relative abundance of Treponema (P < 0.05) and significantly lower relative abundance of Ruminobacter, Ruminococcus, and Streptococcus (P < 0.05). In conclusion, different combinations of sugar and starch concentrations have inconsistent effects on rumen fermentation characteristics, suggesting that the starch in diets cannot be simply replaced with sugar; the combined effects of sugar and starch should be considered to improve the feed utilization rate.

Eye Movements of Spatial Working Memory Encoding in Children with and without Autism: Chunking Processing and Reference Preference.

Individuals with autism spectrum disorder (ASD) experience spatial working memory deficits and show different encoding mechanisms from typical developing (TD) peers. To effectively describe the encoding strategies of those with ASD and highlight their characteristics in cognitive processing, we adopted improved change detection tasks and added eye-movement indicators to investigate the chunking function and reference preference of children with and without ASD. The current study included 20 participants with ASD aged 8-16 and 20 TD children matched for age, gender, and intelligence. Experiment 1 used high/low-structured change detection tasks, and eye-movement indexes were recorded as they memorized the locations of the items to investigate spatial chunking strategies. In Experiment 2, changes in eye movement patterns were observed by adding a frame of reference. The results suggested different encoding strategies in ASD and TD individuals. The ASD group showed local processing bias and had difficulty adopting chunking strategies in spatial working memory. Eye-movement analysis suggested that they rarely showed integrated information processing tendency observed in TD children. Moreover, as a compensatory processing, they were more likely to use the frame of reference. In this study, we compared the spatial chunking strategies and reference preference of children with and without ASD, and eye-movement analysis was used to investigate the processing mechanism. These findings are significant for research on cognitive characteristics of ASD and provide a new focus for working memory training in children with ASD. LAY SUMMARY: The current study suggests that children with autism spectrum disorder are poorer at organizing items into chunks in spatial working memory, but rely more on reference frames. If the purpose of location memory is to strengthen the adaptability of children with autism, it should provide them with more clues or references. If it is for the purpose of intervention such as cognitive training, it should guide them to integrate information to improve the basic cognitive processing efficiency. Autism Res 2021, 14: 897-910. © 2020 International Society for Autism Research and Wiley Periodicals LLC.

Karyopherin α 2 promotes proliferation, migration and invasion through activating NF-κB/p65 signaling pathways in melanoma cells.

AIMS: Melanoma is a fatal malignancy. Karyopherin α 2 (KPNA2) plays an important role in many carcinogenesis. This study was aimed to study the role of KPNA2 in cellular functions and molecular mechanisms of melanoma. MAIN METHODS: We investigated the expression and prognosis of KPNA2 in melanoma using the GEPIA database (http://gepia.cancer-pku.cn/). The effect of KPNA2 on melanoma cells was determined using real-time PCR, western blot, immunofluorescence assay, CCK-8, colony formation, wound healing assay, transwell assay, EMSA, and immunohistochemistry. The influence of KPNA2 on the tumorigenicity of melanoma cells was evaluated in a nude mice model in vivo. KEY FINDINGS: Our results showed that KPNA2 expression is relatively high in melanoma tissues and cells, and melanoma patients with higher expression of KPNA2 had lower overall survival rate and disease free survival rate. KPNA2 promoted proliferation ability and increased the expression of PCNA, Ki67, and C-MYC in melanoma cells. Further, KPNA2 could promote migration and invasion and increase the expression of MMP2 and MMP9. Mechanism studies showed that KPNA2 activated NF-κB/p65 signaling pathways, as evidenced by the nuclear translocation of p65 and increased the expression of COX-2, ICAM-1, iNOS, and MCP1 in melanoma cells. NF-κB inhibitor JSH-23 could reverse the pro-tumor effects of KPNA2 on melanoma cells. Moreover, upregulation of KPNA2 facilitated the tumorigenicity of melanoma cells. SIGNIFICANCE: KPNA2 promotes proliferation, migration and invasion through enhancing NF-κB/p65 signaling pathways in melanoma cells. Our study suggests KPNA2 as a potential therapeutic target for the treatment of melanoma.

Mechanical force promotes the proliferation and extracellular matrix synthesis of human gingival fibroblasts cultured on 3D PLGA scaffolds via TGF­ß expression.

Human gingival fibroblasts (HGFs) are responsible for connective tissue repair and scarring, and are exposed to mechanical forces under physiological and pathological conditions. The exact mechanisms underlying gingival tissue reconstruction under mechanical forces remain unclear. The present study aimfed to investigate the effects of mechanical forces on the proliferation and extracellular matrix synthesis in HGFs by establishing a 3­dimensional (3D) HGF culture model using poly(lactide­co­glycolide) (PLGA) scaffolds. HGFs were cultured in 3D PLGA scaffolds and a mechanical force of 0, 5, 15, 25 or 35 g/cm2 was applied to HGFs for 24 h. A mechanical force of 25 g/cm2 induced the highest proliferation rate, and thus was selected for subsequent experiments. Cell viability was determined using the MTT assay at 0, 24, 48 and 72 h. The expression levels of type I collagen (COL­1) and matrix metallopeptidase (MMP)­1 were examined by reverse transcription­quantitative polymerase chain reaction and ELISA, and transforming growth factor (TGF)­ß expression was evaluated by ELISA. The application of mechanical force on HGFs cultured on the 3D PLGA scaffolds resulted in a significant increase in cell proliferation and COL­1 expression, as well as a decrease in MMP­1 expression. A TGF­ß1 inhibitor was also applied, which attenuated the effects of mechanical force on HGF proliferation, and COL­1 and MMP­1 expression, thus suggesting that TGF­ß signaling pathways may mediate the mechanical force­induced alterations observed in HGFs. In conclusion, these findings helped to clarify the mechanisms underlying mechanical force­induced HGF proliferation and ECM synthesis, which may promote the development of targeted therapeutics to treat various diseases, including gingival atrophy caused by orthodontic treatment.

[Effects of different concentrations of putrescine on proliferation, migration and apoptosis of human skin fibroblasts].

OBJECTIVE: To explore the effects of different concentrations of putrescine on the proliferation, migration and apoptosis of human skin fibroblasts (HSF). METHODS: HSF cultured in the presence of 0.5, 1.0, 5.0, 10, 50, 100, 500, and 1000 µg/ putrescine for 24 h were examined for the changes in the cell proliferation, migration, and apoptosis using MTS assay, Transwell migration assay, and flow cytometry, respectively. RESULTS: Compared with the control cells, HSF cultured with 0.5, 1.0, 5.0, and 10 µg/ putrescine showed significantly increased cell proliferation (P<0.01), and the effect was the most obvious with 1 µg/ putrescine, whereas 500 and 1000 µg/ putrescine significantly reduced the cell proliferation (P<0.01); 50 and 100 µg/ did not obviously affect the cell proliferation (P>0.05). Putrescine at 1 µg/ most significantly enhanced the cell migration (P<0.01), while at higher doses (50, 100, 500, and 1000 µg/) putrescine significantly suppressed the cell migration (P<0.05); 0.5, 5.0, and 10 µg/ putrescine produced no obvious effects on the cell migration (P>0.05). HSF treated with 0.5, 1.0, 5.0, and 10 µg/ putrescine obvious lowered the cell apoptosis rate compared with the control group (P<0.01), and the cell apoptosis rate was the lowest in cells treated with 1 µg/ putrescine; but at the concentrations of 100, 500, and 1000 µg/, putrescine significantly increased the cell apoptosis rate (P<0.01), while 50 µg/ml putrescine produced no obvious effect on cell apoptosis (P>0.05). CONCLUSION: Low concentrations of putrescine can obviously enhance the proliferation ability and maintain normal migration ability of HSF in vitro, but at high concentrations, putrescine can obviously inhibit the cell migration and proliferation and induce cells apoptosis, suggesting the different roles of different concentrations of putrescine in wound healing.

[Biologic effects of different concentrations of putrescine on human umbilical vein endothelial cells].

OBJECTIVE: To explore the effects of different concentrations of putrescine on proliferation, migration, and apoptosis of human umbilical vein endothelial cells (HUVECs). METHODS: HUVECs were routinely cultured in vitro. The 3rd to the 5th passage of HUVECs were used in the following experiments. (1) Cells were divided into 500, 1 000, and 5 000 µg/mL putrescine groups according to the random number table (the same grouping method was used for following grouping), with 3 wells in each group, which were respectively cultured with complete culture solution containing putrescine in the corresponding concentration for 24 h. Morphology of cells was observed by inverted optical microscope. (2) Cells were divided into 0.5, 1.0, 5.0, 10.0, 50.0, 100.0, 500.0, 1 000.0 µg/mL putrescine groups, and control group, with 4 wells in each group. Cells in the putrescine groups were respectively cultured with complete culture solution containing putrescine in the corresponding concentration for 24 h, and cells in control group were cultured with complete culture solution with no additional putrescine for 24 h. Cell proliferation activity (denoted as absorption value) was measured by colorimetry. (3) Cells were divided (with one well in each group) and cultured as in experiment (2), and the migration ability was detected by transwell migration assay. (4) Cells were divided (with one flask in each group) and cultured as in experiment (2), and the cell apoptosis rate was determined by flow cytometer. Data were processed with one-way analysis of variance, Kruskal-Wallis test, and Dunnett test. RESULTS: (1) After 24-h culture, cell attachment was good in 500 µg/mL putrescine group, and no obvious change in the shape was observed; cell attachment was less in 1 000 µg/mL putrescine group and the cells were small and rounded; cells in 5 000 µg/mL putrescine group were in fragmentation without attachment. (2) The absorption values of cells in 0.5, 1.0, 5.0, 10.0, 50.0, 100.0, 500.0, 1 000.0 µg/mL putrescine groups, and control group were respectively 0.588 ± 0.055, 0.857 ± 0.031, 0.707 ± 0.031, 0.662 ± 0.023, 0.450 ± 0.019, 0.415 ± 0.014, 0.359 ± 0.020, 0.204 ± 0.030, and 0.447 ± 0.021, with statistically significant differences among them (χ(2) = 6.86, P = 0.009). The cell proliferation activity in 0.5, 1.0, 5.0, and 10.0 µg/mL putrescine groups was higher than that in control group (P < 0.05 or P < 0.01). The cell proliferation activity in 500.0 and 1 000.0 µg/mL putrescine groups was lower than that in control group (with P values below 0.01). The cell proliferation activity in 50.0 and 100.0 µg/mL putrescine groups was close to that in control group (with P values above 0.05). (3) There were statistically significant differences in the numbers of migrated cells between the putrescine groups and control group (F = 138.662, P < 0.001). The number of migrated cells was more in 1.0, 5.0, and 10.0 µg/mL putrescine groups than in control group (with P value below 0.01). The number of migrated cells was less in 500.0 and 1 000.0 µg/mL putrescine groups than in control group (with P value below 0.01). The number of migrated cells in 0.5, 50.0, and 100.0 µg/mL putrescine groups was close to that in control group (with P values above 0.05). (4) There were statistically significant differences in the apoptosis rate between the putrescine groups and control group (χ(2)=3.971, P=0.046). The cell apoptosis rate was lower in 0.5, 1.0, 5.0, and 10.0 µg/mL putrescine groups than in control group (with P values below 0.05). The cell apoptosis rate was higher in 500.0 and 1 000.0 µg/mL putrescine groups than in control group (with P values below 0.01). The cell apoptosis rates in 50.0 and 100.0 µg/mL putrescine groups were close to the cell apoptosis rate in control group (with P values above 0.05). CONCLUSIONS: Low concentration of putrescine can remarkably enhance the ability of proliferation and migration of HUVECs, while a high concentration of putrescine can obviously inhibit HUVECs proliferation and migration, and it induces apoptosis.