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Abstract Deoxyribonucleotide (DNA) is uniquely programmable and biocompatible, and exhibits unique appeal as a biomaterial as it can be precisely designed and programmed to construct arbitrary shapes. DNA hydrogels are polymer networks comprising cross-linked DNA strands. As DNA hydrogels present programmability, biocompatibility, and stimulus responsiveness, they are extensively explored in the field of biomedicine. In this study, we provide an overview of recent advancements in DNA hydrogel technology. We outline the different design philosophies and methods of DNA hydrogel preparation, discuss its special physicochemical characteristics, and highlight the various uses of DNA hydrogels in biomedical domains, such as drug delivery, biosensing, tissue engineering, and cell culture. Finally, we discuss the current difficulties facing DNA hydrogels and their potential future development. Graphical Abstract

Abstract Background Low concentrations of S100B have neurotrophic effects and can promote nerve growth and repair, which plays an essential role in the pathophysiological and histopathological alterations of major depressive disorder (MDD) during disease development. Studies have shown that plasma S100B levels are altered in patients with MDD. In this study, we investigated whether the plasma S100B levels in MDD differ between genders. Methods We studied 235 healthy controls (HCs) (90 males and 145 females) and 185 MDD patients (65 males and 120 females). Plasma S100B levels were detected via multifactor assay. The Mahalanobis distance method was used to detect the outliers of plasma S100B levels in the HC and MDD groups. The Kolmogorov–Smirnov test was used to test the normality of six groups of S100B samples. The Mann–Whitney test and Scheirer-Ray-Hare test were used for the comparison of S100B between diagnoses and genders, and the presence of a relationship between plasma S100B levels and demographic details or clinical traits was assessed using Spearman correlation analysis. Results All individuals in the HC group had plasma S100B levels that were significantly greater than those in the MDD group. In the MDD group, males presented significantly higher plasma S100B levels than females. In the male group, the plasma S100B levels in the HC group were significantly higher than those in the MDD group, while in the female group, no significant difference was found between the HC and MDD groups. In the male MDD subgroup, there was a positive correlation between plasma S100B levels and years of education. In the female MDD subgroup, there were negative correlations between plasma S100B levels and age and suicidal ideation. Conclusions In summary, plasma S100B levels vary with gender and are decreased in MDD patients, which may be related to pathological alterations in glial cells.

Tissue repair and regeneration have long been a focus of research, with growing evidence highlighting the role played by immunomodulation. The repair of tissue cells can be significantly promoted through the localized immunomodulation of tissues. Hydrogels are increasingly used in this field due to their biocompatibility, versatility, and relative ease of fabrication. This Review highlights the critical role of inflammation and immune development in the complex process of tissue repair after injury, highlighting the dynamic interactions between immune cells that ultimately influence the outcome of tissue healing. We also summarize the precise functions of immune cells and their responses in various tissue injury scenarios, which are critical for harnessing the full potential of immunomodulatory strategies to promote tissue repair. We discuss recent advances in drug-carrying hydrogels, which offer promising avenues for tissue regeneration with precise control of the immune environment for constructive tissue repair. Further advances in our understanding of the mechanisms driving inflammation and immune cell behavior during tissue injury are expected to foster the development of increasingly effective hydrogel-based immunomodulatory strategies to promote tissue repair and regeneration.

Abstract Wound infections pose a major healthcare issue, affecting the well‐being of millions of patients worldwide. Effective intervention and on‐site detection are important in wound management. However, current approaches are hindered by time‐consuming analysis and a lack of technology for real‐time monitoring and prompt therapy delivery. In this study, a smart wound patch system (SWPS) designed for wireless closed‐loop and in‐situ wound management is presented. The SWPS integrates a microfluidic structure, an organic electrochemical transistor (OECT) based sensor, an electrical stimulation module, and a miniaturized flexible printed circuit board (FPCB). The OECT incorporates a bacteria‐responsive DNA hydrogel‐coated gate for continuous monitoring of bacterial virulence at wound sites. Real‐time detection of OECT readings and on‐demand delivery of electrical cues to accelerate wound healing is facilitated by a mobile phone application linked with an FPCB containing low‐power electronics equipped with parallel sensing and stimulation circuitry. In this proof‐of‐concept study, the functionality of the SWPS is validated and its application both in vitro and in vivo is demonstrated. This proposed system expands the arsenal of tools available for effective wound management and enables personalized treatment.

IntroductionProsodic focus marking in Seoul Korean is known to be achieved primarily through prosodic phrasing, different from the use of prosody for this purpose in many other languages. This study investigates how children use prosodic phrasing for focus-marking purposes in Seoul Korean, compared to adults.MethodsUsing a picture-matching game, we elicited semi-spontaneous production of SOV sentences in various focus conditions from monolingual Seoul Korean-speaking children aged 4 to 11 years.ResultsWe found that the children varied prosodic boundaries to distinguish narrow focus from pre-focus and broad focus in a largely adult-like manner at the age of 4 to 5; at this age, they did not distinguish narrow focus from post-focus or contrastive focus using prosodic boundaries, similar to the adults. Their use of the prosodic boundaries in distinguishing the focus conditions was not fully adult-like in terms of frequency until the age of 10 to 11.DiscussionIn conjunction with the findings of previous studies on the acquisition of focus marking in Germanic languages, performed using a similar experimental method, our findings suggest that Seoul Korean-speaking children acquire the use of prosodic phrasing earlier than Dutch-speaking children acquiring the use of pitch accent but slightly later than Stockholm Swedish-speaking children acquiring the use of a prominence-marking high tone. These findings imply that the rate of focus-marking acquisition depends on the transparency of the form-meaning mapping between the phonological cue and focus.

Abstract Mycobacterium tuberculosis (MTB) is the causal pathogen of tuberculosis (TB). Rapid and accurate detection of live MTB is important for transmission control and patient treatment. Here, we described a clustered regularly interspaced short palindromic repeat (CRISPR)‐Cas13a‐based molecular diagnosis approach for rapid and specific detection of live MTB. This detection method, which we termed CRISPR‐Live‐MTB, contained two consecutive reactions including nuclear acid sequence‐based amplification (NASBA) and CRISPR‐Cas13a collateral cleavage reaction. CRISPR‐Live‐MTB could efficiently detect MTB single‐stranded RNA (ssRNA) in 2 hours with high specificity over double‐stranded DNA (dsDNA). Importantly, CRISPR‐Live‐MTB exhibited a limit of detection of 2.4 copies for MTB ssRNA, which was 1000 times lower than that of the clinically used NASBA method. Moreover, lateral flow was integrated into the CRISPR‐Live‐MTB method to enable point‐of‐care testing application with a sensitivity of 95% and a specificity of 100%. Overall, our study demonstrated the feasibility of CRISPR‐Live‐MTB as a rapid, sensitive, and specific approach for live MTB detection.

ABSTRACTLaboratory courses offer a unique opportunity, and sometimes challenge, to engage students in projects where they can experience ownership and authentic science practices. An important science practice is writing, which can lead to increased learning about concepts and science communication. Experiencing a sense of ownership in research can lead to various student outcomes, such as increased motivation, greater interest in research, and higher retention in STEM fields. Although previous work has extracted aspects of ownership from students’ descriptions of research experiences, studies have not examined directly how students define and perceive ownership. In addition, we do not have a clear idea of whether a sense of ownership is related to student attitudes toward scientific writing in a lab course setting. To better understand the relationship between ownership and writing directly from students’ perspectives, we used analysis of student responses to surveys and interviews in an upper-division laboratory course. Using a grounded theory approach for the analysis of 167 survey responses and 9 interviews, we found that students have varying perceptions of project ownership, with the most frequent being opportunities to contribute ideas and shape the project (autonomy), doing the work, and leadership. Students largely perceived that increased ownership had positive influences on their writing, such as increased understanding and thinking, freedom in writing, and increased motivation. Learning about how students perceive ownership in the context of a lab course is useful for considering how lab course structure may support the development of a sense of ownership and may influence how we can engage students in meaningful writing practices.

Abstract Background Workplace violence is a worldwide concern, and particularly affects nursing students. It has a seriously negative impact on nursing students’ clinical learning experience and their physical and mental health. This study explored whether there are differences in psychological responses and coping styles among different gender nursing students after exposure to workplace violence, and investigated the causes for these differences. Methods We enrolled 22 nursing undergraduates from Guangzhou Medical University and Zunyi Medical University, China. Phenomenological qualitative research and online semi-structured interviews were conducted. The data were analyzed by the Colaizzi seven-step content analysis method. Results Two categories were collated: psychological experience and coping styles. Three themes of the former were extracted: negative emotional experience, low level of professional identity, and negative effect on self-efficacy. Two themes of the latter: responses to violence and adjustment after violence. In addition, fourteen subthemes were extracted. Conclusions Different gender nursing students have different psychological experience and coping styles in the face of workplace violence. The causes of the differences are likely related to sociocultural factors and psychological gender status.

Background: Mitochondrial redox imbalance underlies the pathophysiology of type2 diabetes mellitus (T2DM), and is closely related to tissue damage and dysfunction. Studies have shown the beneficial effects of dietary strategies that elevate β-hydroxybutyrate (BHB) levels in alleviating T2DM. Nevertheless, the role of BHB has not been clearly elucidated. Methods: We performed a spectral study to visualize the preventive effects of BHB on blood and multiorgan mitochondrial redox imbalance in T2DM mice via using label-free resonance Raman spectroscopy (RRS), and further explored the impact of BHB therapy on the pathology of T2DM mice by histological and biochemical analyses. Findings: Our data revealed that RRS-based mitochondrial redox states assay enabled clear and reliable identification of the improvement of mitochondrial redox imbalance by BHB, evidenced by the reduction of Raman peak intensity at 750 cm−1, 1128 cm−1 and 1585 cm−1 in blood, tissue as well as purified mitochondria of db/db mice and the increase of tissue mitochondrial succinic dehydrogenase (SDH) staining after BHB treatment. Exogenous supplementation of BHB was also found to attenuate T2DM pathology related to mitochondrial redox states, involving organ injury, blood glucose control, insulin resistance and systemic inflammation. Interpretation: Our findings provide strong evidence for BHB as a potential therapeutic strategy targeting mitochondria for T2DM.

DNMT3A participates in de novo methylation, yet its impact on the proliferation of testicular Sertoli cells remains unclear. Development-specific methylation has been proven to be associated with cellular development. Therefore, in this study, we simulated DNMT3A expression pattern during testicular development by DNMT3A interference. Then, RRBS and RNA-seq were used to decipher DNMT3A regulatory mechanisms on Sertoli cell proliferation. Immunofluorescence staining revealed the expression of DNMT3A in the Sertoli cells of the prepubertal testis. DNMT3A was demonstrated to inhibit the cell cycle and proliferation of Sertoli cells, while promoting cell apoptosis. After transfected with DNMT3A interference, a total of 560 DEGs and 2,091 DMGs produced by DNMT3A interference were identified between two treated groups, respectively. Integrating the results from RRBS and RNA-seq, the overlapping genes between DMGs and DEGs were found to be enriched in the Gene Ontology (GO) terms related to cellular development and the Apelin signaling pathway. The present study demonstrated the impact of DNMT3A on the proliferation of porcine testicular Sertoli cells, suggesting that DNMT3A primarily acts through the Apelin signaling pathway. These findings provide valuable insights into how DNMT3A influences testicular development and health, offering new perspectives.

Normal testicular development ensures the process of spermatogenesis, which is a complex biological process. The sustained high productivity of spermatogenesis throughout life is predominantly attributable to the constant proliferation and differentiation of spermatogonial stem cells (SSCs). The self-renewal and differentiation processes of SSCs are strictly regulated by the SSC niche. Therefore, understanding the developmental pattern of SSCs is crucial for spermatogenesis. The Shaziling pig is a medium-sized indigenous pig breed originating from central China. It is renowned for its superior meat quality and early male sexual maturity. The spermatogenic ability of the boars is of great economic importance to the pig industry. To investigate testicular development, particularly the pattern of SSC development in Shaziling pigs, we used single-cell transcriptomics to identify gene expression patterns in 82,027 individual cells from nine Shaziling pig testes at three key postnatal developmental stages. We generated an unbiased cell developmental atlas of Shaziling pig testicular tissues. We elucidated the complex processes involved in the development of SSCs within their niche in the Shaziling pig. Specifically, we identified potential marker genes and cellular signaling pathways that regulate SSC self-renewal and maintenance. Additionally, we proposed potential novel marker genes for SSCs that could be used for SSC isolation and sorting in Shaziling pigs. Furthermore, by immunofluorescence staining of testicular tissues of different developmental ages using marker proteins (UCHL1 and KIT), the developmental pattern of the spermatogonia of Shaziling pigs was intensively studied. Our research enhances the comprehension of the development of SSCs and provides a valuable reference for breeding Shaziling pigs.

The role of lncRNA and circRNA in wheat grain development is still unclear. The objectives of this study were to characterize the lncRNA and circRNA in the wheat grain development and to construct the interaction network among lncRNA, circRNA, and their target miRNA to propose a lncRNA–circRNA–miRNA module related to wheat grain development. Full transcriptome sequencing on two wheat varieties (Annong 0942 and Anke 2005) with significant differences in 1000-grain weight at 10 d (days after pollination), 20 d, and 30 d of grain development were conducted. We detected 650, 736, and 609 differentially expressed lncRNA genes, and 769, 1054, and 1062 differentially expressed circRNA genes in the grains of 10 days, 20 days and 30 days after pollination between Annong 0942 and Anke 2005, respectively. An analysis of the lncRNA–miRNA and circRNA–miRNA targeting networks reveals that circRNAs exhibit a more complex and extensive interaction network in the development of cereal grains and the formation of grain shape. Central to these interactions are tae-miR1177, tae-miR1128, and tae-miR1130b-3p. In contrast, lncRNA genes only form a singular network centered around tae-miR1133 and tae-miR5175-5p when comparing between varieties. Further analysis is conducted on the underlying genes of all target miRNAs, we identified TaNF-YB1 targeted by tae-miR1122a and TaTGW-7B targeted by miR1130a as two pivotal regulatory genes in the development of wheat grains. The quantitative real-time PCR (qRT-PCR) and dual-luciferase reporter assays confirmed the target regulatory relationships between miR1130a-TaTGW-7B and miR1122a-TaNF-YB1. We propose a network of circRNA and miRNA-mediated gene regulation in the development of wheat grains.

Laboratory courses offer a unique opportunity, and sometimes challenge, to engage students in projects where they can experience ownership and authentic science practices. An important science practice is writing, which can lead to increased learning about concepts and science communication. Experiencing a sense of ownership in research can lead to various student outcomes, such as increased motivation, greater interest in research, and higher retention in STEM fields. Although previous work has extracted aspects of ownership from students' descriptions of research experiences, studies have not examined directly how students define and perceive ownership. In addition, we do not have a clear idea of whether a sense of ownership is related to student attitudes toward scientific writing in a lab course setting. To better understand the relationship between ownership and writing directly from students' perspectives, we used analysis of student responses to surveys and interviews in an upper-division laboratory course. Using a grounded theory approach for the analysis of 167 survey responses and 9 interviews, we found that students have varying perceptions of project ownership, with the most frequent being opportunities to contribute ideas and shape the project (autonomy), doing the work, and leadership. Students largely perceived that increased ownership had positive influences on their writing, such as increased understanding and thinking, freedom in writing, and increased motivation. Learning about how students perceive ownership in the context of a lab course is useful for considering how lab course structure may support the development of a sense of ownership and may influence how we can engage students in meaningful writing practices.

Estrus involves a series of complex physiological signs and changes in behavior before ovulation, which play a crucial role in animal reproduction. However, there have been few studies that evaluate behaviors during the different stages of estrus cycle in female rabbits. Therefore, more detailed information is needed on distinguishing the various stages of the estrous cycle. This study explored the behavioral and physiological differences at various estrous cycle stages in female New Zealand White rabbits. The continuous recording method was employed to record the daily behaviors of twenty postpartum female rabbits during the estrous cycle. Compared with the diestrus stage, the duration of foraging and drinking behavior in estrus decreased significantly, and the frequency of grooming and biting behaviors increased (p < 0.05). Differences in reproductive hormone levels (FSH, LH, P4, and E2) and follicle development were measured at each stage via ELISA and HE staining. The FSH and LH levels showed an increasing trend and then decreased, with the lowest being in late estrus (p < 0.05). The P4 level was the lowest in estrus (p < 0.05), and E2 showed a gradually increasing trend. There was no significant difference in the number of primordial follicles at each stage, but the number of primary follicles in estrus was significantly higher than at the other stages (p < 0.05). To further understand the molecular regulation mechanism of the estrous cycle in female rabbits, we analyzed the ovarian transcription patterns of female rabbits in diestrus (D group) and estrus (E group) employing RNA-seq. A total of 967 differentially expressed genes (DEGs) were screened from the ovaries of female rabbits between the diestrus and estrus groups. A KEGG analysis of DEGs enriched in the estrogen signaling pathway, aldosterone synthesis, and secretion pathway, such as CYP19A1 and IGF1R, was performed. The rabbits’ behavior, related physiological hormones, and molecular regulation also differed at different estrous cycle stages. The results provide recommendations for the adequate management practices of postpartum re-estrus and breeding female rabbits.

Magnetic MgxCoyZn(1-x-y)Fe2O4 nanoparticles were successfully prepared by the rapid combustion approach, and SEM, XRD, VSM, EDX, and FTIR techniques were applied for their characterization. The influence of the element ratios (Mg2+, Co2+, and Zn2+) in magnetic MgxCoyZn(1-x-y)Fe2O4 nanoparticles on their properties was explored. To acquire a larger specific surface area for better adsorption of methyl blue (MB), magnetic Mg0.4Co0.5Zn0.1Fe2O4 nanoparticles calcined at 400°C for 2 h with 25 mL anhydrous ethanol were selected, and their average particle size and the saturation magnetization were about 81.3 nm and 13.5 emu·g-1, respectively. Adsorption kinetics models and adsorption isotherm models were applied to research the adsorption characteristics of MB onto magnetic Mg0.4Co0.5Zn0.1Fe2O4 nanoparticles. The pseudo-second-order kinetics model (R2>0.99) and Temkin isotherm model (R2=0.9887) were the most consistent with the data, indicating that the adsorption was the chemical multilayer adsorption mechanism, and the process was an exothermic reaction. The E of the Dubinin-Radushkevich (D-R) isotherm model was 0.2347 KJ·mol-1, indicating the adsorption involved physical adsorption besides chemical adsorption. The ΔG0 and ΔH0 (ΔH0=−10.38 KJ·mol-1) of the adsorption process of MB adsorbed onto magnetic Mg0.4Co0.5Zn0.1Fe2O4 nanoparticles measured through the thermodynamic experiment were both less than 0, which proved that the process was a spontaneous exothermic reaction. The adsorption capacity of MB onto magnetic Mg0.4Co0.5Zn0.1Fe2O4 nanoparticles increased with the pH of MB solution increasing from 2 to 4 at room temperature, and it had no significant change when the pH of MB solution was 4-12, while the relative removal rate was 98.75% of the first one after 2 cycles. The electrochemical impedance spectroscopy (EIS) and the cyclic voltammetry (CV) data further demonstrated that MB was adsorbed onto magnetic Mg0.4Co0.5Zn0.1Fe2O4 nanoparticles.

Background: Major depressive disorder (MDD) is a severe mental disorder characterized by reduced gray matter volume (GMV). To date, the pathogenesis of MDD remains unclear, but neurotrophic factors play an essential role in the pathophysiological alterations of MDD during disease development. In particular, plasma glial cell line-derived neurotrophic factor (GDNF) has been suggested as a potential biomarker that may be associated with disease activity and neurological progression in MDD. Our study investigated whether plasma GDNF levels in MDD patients and healthy controls (HCs) are correlated with GMV alterations. Methods: We studied 54 MDD patients and 48 HCs. The effect of different diagnoses on whole-brain GMV was investigated using ANOVA (Analysis of Variance). The threshold of significance was p

RNA N6-methyladenosine (m6A) modification is one of the principal post-transcriptional modifications and plays a dynamic role in testicular development and spermatogenesis. However, the role of m6A in porcine testis is understudied. Here, we performed a comprehensive analysis of the m6A transcriptome-wide profile in Shaziling pig testes at birth, puberty, and maturity. We analyzed the total transcriptome m6A profile and found that the m6A patterns were highly distinct in terms of the modification of the transcriptomes during porcine testis development. We found that key m6A methylated genes (AURKC, OVOL, SOX8, ACVR2A, and SPATA46) were highly enriched during spermatogenesis and identified in spermatogenesis-related KEGG pathways, including Wnt, cAMP, mTOR, AMPK, PI3K-Akt, and spliceosome. Our findings indicated that m6A methylations are involved in the complex yet well-organized post-transcriptional regulation of porcine testicular development and spermatogenesis. We found that the m6A eraser ALKBH5 negatively regulated the proliferation of immature porcine Sertoli cells. Furthermore, we proposed a novel mechanism of m6A modification during testicular development: ALKBH5 regulated the RNA methylation level and gene expression of SOX9 mRNA. In addition to serving as a potential target for improving boar reproduction, our findings contributed to the further understanding of the regulation of m6A modifications in male reproduction.

The functions of non-agricultural crops unearthed from archaeological sites mainly pertain to their usage as livestock feed. However, the studies of livestock feed have predominantly relied on qualitative analysis, which often lacks descriptive objectivity and relies heavily on subjective feelings and experiences. In this paper, we aim to address this gap by focusing on quantitative analysis, utilizing macro-plant remains from the data of seventy-five archaeological settlements and one archaeological investigation in northern China spanning the Neolithic Age to the Bronze Age, as well as stable isotope analyses of carbon and nitrogen. This research delves into various aspects, including the exploration of the plant resources and livestock farming and the categorization of feed types for cattle and pigs in captivity. By employing quantitative analysis, we can gain a more comprehensive and objective understanding of these subjects. This approach aligns with studies on ancient livestock management and feed diversity. In essence, the discussion of civilization development and social changes during the Pre-Qin period holds significant value when considering forage analysis, just as crop analysis has proven insightful. By focusing on the quantitative analysis of non-agricultural crops and their role as livestock feed, we can shed light on important aspects of ancient societies and their agricultural practices.

ZnV2O6 modified by graphite is a promising photocatalyst to improve CO2 reduction efficiency. In this paper, the density functional theory studied the energy band structure, charge transfer, and optical properties of graphene/ZnV2O6(001) heterojunction. The mechanism of performance enhancement of graphene/ZnV2O6(001) heterojunction is explored. The results state that the energy band structure of graphene/ZnV2O6(001) is reduced to 0.025eV due to graphene's hybridization. Under visible light irradiation, electrons transition from the top of the ZnV2O6 valence band to the conduction band and are excited to graphene due to the electrostatic field, which promotes electron–hole pairs' separation inhibit the recombination of carriers. The ground state interface electron transfer encourages the increase of surface electrons of graphene and accumulates a large number of holes on ZnV2O6(001). A good combination of ZnV2O6(001) and graphene is beneficial to charge transfer and improve photocatalytic efficiency. The calculated results show that the band structure of graphene/ZnV2O6(001) heterojunction has apparent advantages in improving the visible light photocatalytic efficiency, consistent with the present experimental observation. Besides, our work can also serve as the basis for designing vanadate-based semiconductor heterostructures in photocatalysts and solar cells.

The purpose of this experiment was to investigate the effects of different sources of calcium on immune performance, diarrhea rate, intestinal barrier, and post-intestinal flora structure and function in weaned piglets. A total of 1,000 weaned piglets were randomly assigned to five groups 10 replicate pens per treatment, 20 piglets per pen and fed calcium carbonate, calcium citrate, multiple calcium, and organic trace minerals of different concentrations of acidifier diets. The results of the study showed that the replacement of calcium carbonate with calcium citrate and multiple calcium had almost no significant effect on immune indexes (IL-1β, IL-6, IL-10, TNF-α) of piglets compared with the control group (p > 0.05). The five groups did not show a change in the diarrhea rate and diarrhea index (p > 0.05). The diet containing multiple calcium dramatically decreased the TP compared to the C and L diet (p < 0.05). No significant difference in HDL was noted in the five groups (p > 0.05). However, the concentration of LDL in blood in the multiple calcium group was significantly higher than that in groups L and D (p < 0.05). Moreover, the concentration of Glu in blood in the multiple calcium group was significantly higher than that in group C (p < 0.05). Compared with the control group, calcium citrate plus organic trace minerals diet markedly increased UCG-005 abundance in the colon (p < 0.05). In addition, the relative abundance of Prevotellaceae_NK3B31_group had an upward trend in the colon of the M group compared to the D group (p = 0.070). Meanwhile, calcium citrate plus organic trace minerals diet markedly increased Clostridium_sensu_stricto_1 abundance in the colon (p < 0.05). Metagenomic predictions by PICRUSt suggested that the colonic and fecal microbiota was mainly involved in carbohydrate metabolism, amino acid metabolism, energy metabolism, and metabolism of cofactors and vitamins.

The purpose of this experiment was to investigate the effects of different protein levels on lipid metabolism and gut microbes in mice of different genders. A total of 60 mice (30 female and 30 male) were randomly assigned to six groups and fed female mice with low protein diet (FLP), basal protein diet (FBD), and high protein diet (FHP). Similarly, the male mice fed with low protein diet (MLP), basal protein diet (MBD), and high protein diet (MHP). The low protein diet contained 14% CP, the basal diet contained 20% CP, and the high protein diet contained 26% CP. The results of the study showed that both basal and high protein diets significantly reduced the perirenal adipose tissues (PEAT) index in male mice compared to low protein diet (p < 0.05). For the gut, the FHP significantly increased the relative gut weight compared to the FBD and FLP (p < 0.05). At the same time, the FHP also significantly increased the relative gut length compared with the FBD and FLP (p < 0.05). The MHP significantly increased TC concentration compared with the MLP (p < 0.05), and the MBD tended to increase TC concentration compared with the MLP in serum (p = 0.084). The histomorphology result of the jejunum and ileum showed that a low protein diet was beneficial to the digestion and absorption of nutrients in the small intestine of mice. While different protein levels had no effect on the total number of fecal microbial species in mice, different protein levels had a significant effect on certain fecal microbes in mice, the absolute abundance of Verrucomicrobia in the feces of male mice was significantly higher in both high and basal protein diets than in the low protein diet (p < 0.05). The high protein diet significantly reduced the absolute abundance of Patescibacteria in the feces of female mice compared to both the basal and low protein diets (p < 0.05). The absolute abundance of Patescibacteria in male feces was not affected by dietary protein levels (p > 0.05). Taken together, our results suggest that a low protein diet can alter fat deposition and lipid metabolism in mice, and that it benefited small intestinal epithelial structure and microbes.

Despite a well-documented effect of calcium on the piglet's intestinal microbiota composition, it is less known about changes in microbial function or the effect of different sources of calcium. The experiment was designed to study the effects of dietary calcium from different sources on production, immune indexes, antioxidant capacity, serum biochemical indexes, and intestinal microflora of weaning piglets. A total of 1,000 piglets were randomly assigned to five groups (10 replicate pens per treatment with 20 pigs per pen) and fed diets supplemented with calcium carbonate, calcium citrate, multiple calcium, organic trace minerals, and different concentrations of acidifier. The results showed that the replacement of calcium carbonate with calcium citrate and multiple calcium had almost no significant difference in the growth performance of pigs compared with the control group, and only the diet of multiple calcium dramatically decreased the average daily feed intake (ADFI) compared to the calcium citrate diet on days 15–28 (p < 0.05). The five groups did not change the content of MDA, SOD, and GSH-Px (p > 0.10). A similar situation occurs in the immune function of the blood. There was no significant effect in immune indexes (IgA, IgG, and IgM) among different treatments after weaning at 6 weeks for piglets (p > 0.10). The 16S rRNA sequencing of ileal and cecal microbiota revealed that only the relative abundance of Actinobacteriota at the phyla level was significantly greater in the ileum of the A group compared to the other treatments (p < 0.05). There was a clear effect on seven bacteria in the top 30 genera of ileum and cecum for five groups (p < 0.05). The result of PICRUSt predicted that the intestinal microbe was mainly involved in carbohydrate and amino acid metabolism, membrane transport, and metabolism of cofactors and vitamins. Besides, adding calcium citrate to a weaned piglet diet is better than other choices from the third week to the fourth week. In conclusion, diets with different calcium sources changed ADFI and some intestinal microbial composition of weaned piglets but had little effect on intestinal microbial function.

The regulatory role of non-CpG methylation in mammals has been important in whole-genome bisulfite sequencing. It has also been suggested that non-CpG methylation regulates gene expression to affect the development and health of mammals. However, the dynamic regulatory mechanisms of genome-wide, non-CpG methylation during testicular development still require intensive study. In this study, we analyzed the dataset from the whole-genome bisulfite sequencing (WGBS) and the RNA-seq of precocious porcine testicular tissues across two developmental stages (1 and 75 days old) in order to explore the regulatory roles of non-CpG methylation. Our results showed that genes regulated by non-CpG methylation affect the development of testes in multiple pathways. Furthermore, several hub genes that are regulated by non-CpG methylation during testicular development—such as VEGFA, PECAM1, and FZD7—were also identified. We also found that the relative expression of FZD7 was downregulated by the zebularine-induced demethylation of the first exon of FZD7. This regulatory relationship was consistent with the results of the WGBS and RNA-seq analysis. The immature porcine Sertoli cells were transfected with RNAi to mimic the expression patterns of FZD7 during testicular development. The results of the simulation test showed that cell proliferation was significantly impeded and that cell cycle arrest at the G2 phase was caused by the siRNA-induced FZD7 inhibition. We also found that the percentage of early apoptotic Sertoli cells was decreased by transfecting them with the RNAi for FZD7. This indicates that FZD7 is an important factor in linking the proliferation and apoptosis of Sertoli cells. We further demonstrated that Sertoli cells that were treated with the medium collected from apoptotic cells could stimulate proliferation. These findings will contribute to the exploration of the regulatory mechanisms of non-CpG methylation in testicular development and of the relationship between the proliferation and apoptosis of normal somatic cells.

Abstract Large‐scale screening of molecules heavily relies on phenotyping of small living organisms during preclinical development. However, deep profiling candidate therapeutics on whole animals typically requires laborious manipulations and anesthetic treatment using traditional techniques or automated tools. Here, a novel fish capsule system that combines automated zebrafish encapsulating technology and droplet microarray strategy for in vivo functional screening of mono/polytherapies is described. This platform enables automated, rapid zebrafish orientation and immobilization in agarose to generate large‐scale fish capsules by using a microfluidic device. Based on the effect of discontinuous dewetting, the prompt trapping of fish capsules in the aqueous arrays is successfully demonstrate. This system provides the capability to integrate pharmaceutical treatments with real‐time multispectral microscopic imaging in a simple, pipetting‐free and highly parallel manner. Coupling with machine learning algorithms, a small library of compounds is screened and analyzed, and clues about how to exploit compound combinations as therapeutic candidates are obtained. It is believed that this proposed strategy can be readily applied to multiple fields and is especially useful in the exploration of combinatorial drugs with limited amounts of samples and resources to accelerate the identification of novel therapeutics for precision medicines.

Exploring the mechanism of cadmium (Cd) accumulation in Cd-safe rice lines is beneficial for ensuring rice safety. D62B, a Cd-safe rice line, accumulates less than 0.2 mg Cd kg-1 in the brown rice due to strong capacity of Cd retention in the roots, and the root cell wall (RCW) polysaccharides play important roles. However, specific underlying mechanism of Cd binding on the polysaccharides is little known. In this study, the role of polysaccharides, especially pectin and hemicellulose 1 (HC1), in RCW of D62B was investigated by adsorption experiments and Fourier Transform Infrared Spectroscopy (FTIR) analysis compared with a common rice line (Luhui17). Cadmium was adsorbed on RCW of two rice lines by a multilayer and inhomogeneous chemisorption way with the force of ion transfer or exchange. Cadmium was adsorbed on RCW rapidly at first stage with the limit of internal and external diffusion, and gradually reached saturation. With the removal of pectin, the Cd adsorption rate, maximum Cd adsorption amount and the shift degree of carboxyl groups in the RCW of D62B sharply decreased, which showed advantages compared with Luhui17. Sequential removal of HC1 further decreased the maximum Cd adsorption amount and the shift degree of hydroxyl groups. The results showed that more available functional groups, especially carboxyl groups in pectin and hydroxyl groups in HC1, contributed to Cd immobilization within the RCW of Cd-safe rice line, thus limiting Cd translocation to the shoot and reducing Cd accumulation in the brown rice.

MicroRNAs (miRNAs) have been widely identified in porcine testicular tissues and implicated as crucial regulators of proliferation, apoptosis, and differentiation in porcine spermatogenesis related cells. However, the function roles of most of the miRNAs that have been identified in Sertoli cells are poorly understood. In the present study, six experiments were conducted to study the regulatory role of miR-10b in porcine immature Sertoli cells. In experiment 1, the results showed that the relative mRNA expression level of miR-10b in porcine testicular tissues decreased quadratically (P0.05) on porcine immature Sertoli cell apoptosis. In experiment 6, the co-transfection treatment results showed that miR-10b promoted (P

Sertoli cells are central and essential coordinators of spermatogenesis. Accumulating evidence has demonstrated that miRNAs participate in the regulation of Sertoli cell growth. However, the functions and the regulatory mechanisms of miRNAs in Sertoli cells of domestic animals remain largely unknown. Here we report that miR-222 overexpression repressed cell cycle progression and proliferation and promoted the apoptosis of immature porcine Sertoli cells, whereas miR-222 inhibition resulted in the opposite result. miR-222 directly targeted the 3′-UTR of the GRB10 gene and inhibited its mRNA abundance. An siRNA-induced GRB10 knockdown showed similar effects as did miR-222 overexpression on cell proliferation and apoptosis and further attenuated the role of miR-222 inhibition. Furthermore, both miR-222 overexpression and GRB10 inhibition repressed the phosphorylation of PI3K and AKT, the key elements of the PI3K/AKT signaling pathway, whereas GRB10 inhibition offsets the effects of the miR-222 knockdown. Overall, we concluded that miR-222 suppresses immature porcine Sertoli cell growth by targeting the GRB10 gene through inactivation of the PI3K/AKT signaling pathway. This study provides novel insights into the epigenetic regulation of porcine spermatogenesis by determining the fate of Sertoli cells.

Background/Aims: Glioblastoma is the most common and aggressive brain tumor and carries a poor prognosis. Previously, we found that neurotensin receptor 1 (NTSR1) contributes to glioma progression, but the underlying mechanisms of NTSR1 in glioblastoma invasion remain to be clarified. The aim of this study was to investigate the molecular mechanisms of NTSR1 in glioblastoma invasion. Methods: Cell migration and invasion were evaluated using wound-healing and transwell assays. Cell proliferation was detected using CCK-8. The expression of NTSR1, Jun, and suppressor of cytokine signaling 6 (SOCS6) was detected using western blotting. The expression of miR-494 was detected by Quantitative real-time PCR. Chromatin immunoprecipitation assay was performed to examine the interaction between Jun and miR-494 promoter. Dual-luciferase reporter assay and western blotting were performed to identify the direct regulation of SOCS6 by miR-494. An orthotopic xenograft mouse model was conducted to assess tumor growth and invasion. Results: NTSR1 knockdown attenuated the invasion of glioblastoma cells. Jun was positively regulated by NTSR1, which promoted miR-494 expression through binding to miR-494 promoter. SOCS6 was confirmed as a direct target of miR-494, thus, NTSR1-induced miR-494 upregulation resulted in SOCS6 downregulation. Both miR-494 and SOCS6 were involved in the NTSR1-induced invasion of glioblastoma cells. In vivo, tumor invasion and growth were inhibited by NTSR1 knockdown, but were restored with miR-494 overexpression. Conclusion: NTSR1 knockdown inhibited glioblastoma invasion via the Jun/miR-494/SOCS6 axis.

The quantity of Sertoli cells in the adult testis decides the daily gamete formation, and accumulating evidence indicates that epigenetic factors regulate the proliferation of Sertoli cells. Research on the function and regulatory mechanism of microRNAs (miRNAs) in Sertoli cells has not been comprehensive yet, especially on domestic animals. In this article, we report that miR-126 controls the proliferation and apoptosis of immature porcine Sertoli cells based on previous studies. Our results confirmed that miR-126 elevation promotes cell cycle progression, cell proliferation and represses cell apoptosis; on the contrary, the inhibitory effects of miR-126 result in the opposite. The phosphoinositide-3-kinase regulatory subunit 2 (PIK3R2) gene, a member of the PI3K family, was verified as a direct target of miR-126 using the dual-luciferase reporter analysis. miR-126 negatively regulated the mRNA and protein expression level of PIK3R2 in immature porcine Sertoli cells. siRNA-induced PIK3R2 inhibition caused similar effects as miR-126 overexpression and eliminated the influences of miR-126 knockdown in immature porcine Sertoli cells. In addition, both miR-126 overexpression and PIK3R2 inhibition elevated the phosphorylation of PI3K and AKT, whereas the miR-126 knockdown demonstrated the contrary result. In short, miR-126 controls the proliferation and apoptosis of immature porcine Sertoli cells by targeting the PIK3R2 gene through the PI3K/AKT signaling pathway. The research supplies a theoretical and practical foundation for exploring the functional parts of miR-126 in swine sperm by defining the destiny of immature Sertoli cells.

The integrated stress response (ISR), a pivotal protein homeostasis network, plays a critical role in the formation of long-term memory (LTM). The precise mechanism by which the ISR controls LTM is not well understood. Here, we report insights into how the ISR modulates the mnemonic process by using targeted deletion of the activating transcription factor 4 (ATF4), a key downstream effector of the ISR, in various neuronal and non-neuronal cell types. We found that the removal of ATF4 from forebrain excitatory neurons (but not from inhibitory neurons, cholinergic neurons, or astrocytes) enhances LTM formation. Furthermore, the deletion of ATF4 in excitatory neurons lowers the threshold for the induction of long-term potentiation, a cellular model for LTM. Transcriptomic and proteomic analyses revealed that ATF4 deletion in excitatory neurons leads to upregulation of components of oxidative phosphorylation pathways, which are critical for ATP production. Thus, we conclude that ATF4 functions as a memory repressor selectively within excitatory neurons. [ABSTRACT FROM AUTHOR]

Sepsis is a life-threatening infectious disease. Mitochondrial dysfunction is widespread in severe sepsis. The myocardium contains a large number of mitochondria, and the survival rate of sepsis decreases sharply when cardiac dysfunction is involved. Vericiguat (BAY 1021189) is a novel drug for the prevention of heart failure. In this study, we evaluated the mitochondrial function of septic mice and drug-treated mice by resonance Raman spectroscopy (RRS). RRS is a non-invasive and label-free technique that can identify molecular vibrations by their unique fingerprints, making it ideal for quantitative studies. By choosing 532 nm as the excitation wavelength, which is close to the absorption peak of cytochrome, we can greatly enhance the Raman signal of mitochondrial redox state. RRS can accurately identify the Raman characteristic peak at 750 cm-1, 1128 cm-1 and 1585 cm-1 attributed to the reduced cytochrome in septic mice. We found that the intensity of the characteristic peak was significantly decreased in septic mice, indicating an imbalance of mitochondrial redox function, while the function was improved in the drug-treated group. It proves that BAY has the potential as a novel treatment for mitochondrial dysfunction in sepsis. [ABSTRACT FROM AUTHOR]