Effects of exogenous glycine betaine on growth and development of tomato seedlings under cold stress.

Low temperature is a type of abiotic stress affecting the tomato (Solanum lycopersicum) growth. Understanding the mechanisms and utilization of exogenous substances underlying plant tolerance to cold stress would lay the foundation for improving temperature resilience in this important crop. Our study is aiming to investigate the effect of exogenous glycine betaine (GB) on tomato seedlings to increase tolerance to low temperatures. By treating tomato seedlings with exogenous GB under low temperature stress, we found that 30 mmol/L exogenous GB can significantly improve the cold tolerance of tomato seedlings. Exogenous GB can influence the enzyme activity of antioxidant defense system and ROS levels in tomato leaves. The seedlings with GB treatment presented higher Fv/Fm value and photochemical activity under cold stress compared with the control. Moreover, analysis of high-throughput plant phenotyping of tomato seedlings also supported that exogenous GB can protect the photosynthetic system of tomato seedlings under cold stress. In addition, we proved that exogenous GB significantly increased the content of endogenous abscisic acid (ABA) and decreased endogenous gibberellin (GA) levels, which protected tomatoes from low temperatures. Meanwhile, transcriptional analysis showed that GB regulated the expression of genes involved in antioxidant capacity, calcium signaling, photosynthesis activity, energy metabolism-related and low temperature pathway-related genes in tomato plants. In conclusion, our findings indicated that exogenous GB, as a cryoprotectant, can enhance plant tolerance to low temperature by improving the antioxidant system, photosynthetic system, hormone signaling, and cold response pathway and so on.

Association of changes in expression of HDAC and SIRT genes after drug treatment with cancer cell line sensitivity to kinase inhibitors.

Histone deacetylases (HDACs) and sirtuins (SIRTs) are important epigenetic regulators of cancer pathways. There is a limited understanding of how transcriptional regulation of their genes is affected by chemotherapeutic agents, and how such transcriptional changes affect tumour sensitivity to drug treatment. We investigated the concerted transcriptional response of HDAC and SIRT genes to 15 approved antitumor agents in the NCI-60 cancer cell line panel. Antitumor agents with diverse mechanisms of action induced upregulation or downregulation of multiple HDAC and SIRT genes. HDAC5 was upregulated by dasatinib and erlotinib in the majority of the cell lines. Tumour cell line sensitivity to kinase inhibitors was associated with upregulation of HDAC5, HDAC1, and several SIRT genes. We confirmed changes in HDAC and SIRT expression in independent datasets. We also experimentally validated the upregulation of HDAC5 mRNA and protein expression by dasatinib in the highly sensitive IGROV1 cell line. HDAC5 was not upregulated in the UACC-257 cell line resistant to dasatinib. The effects of cancer drug treatment on expression of HDAC and SIRT genes may influence chemosensitivity and may need to be considered during chemotherapy.

Insight into interactions between microplastics and fulvic acid: Mechanisms affected by microplastics type.

Microplastics (MPs) can interact with dissolved organic matter (DOM), a common component found in the environment. However, the effect of MPs type on its interaction with DOM has not been systematically studied. Therefore, the binding properties of different MPs with fulvic acid (FA) were explored in this study. The results showed that polypropylene (PP) and polyethylene (PE) had higher adsorption affinity for FA than polystyrene (PS) and polyvinyl chloride (PVC). The interaction between MPs and FA conformed to the pseudo-first-order model and Freundlich model (except PS). The interaction mechanisms between various MPs tested in this paper and FA are considered to be different. PP, PE and PS interacted with the aromatic structure of FA and were entrapped in the FA polymers by the carboxyl groups and CO bonds, resulting in a highly conjugated co-polymer, suggesting that oxygen-containing functional groups played a key role. However, it was assumed that the interaction between PVC and FA was more likely to be caused by hydrophobic interaction. This research will help to enhance our comprehension of the environmental behavior of MPs and their interaction with the DOM specifically.

The impact of the COVID-19 pandemic on the trend of ventilator associated pneumonia incidence in a non-COVID-19 dedicated hospital.

INTRODUCTION: Ventilator associated pneumonia (VAP) is one of the most common hospital-acquired infections for intensive care units in China. Since the COVID-19 outbreak, "Lockdown Wuhan" and other infection control strategies had been implemented in China. The impact of the policies on VAP prevention was estimated in a non-COVID-19 dedicated hospital. METHODOLOGY: We analyzed the VAP trends of 6 intensive care units in a non-COVID-19 dedicated hospital from 2018 to 2020 by Joinpoint regression analysis. The information related to infected VAP patients, VAP surveillance were retrieved from an active surveillance system. RESULTS: There was an obvious decrease in the overall admissions and inpatients of ICUs since January 2020. The overall incidence of VAP was 6.1 episodes per 1000 IMV days. The 30-day case fatality was 16.8%. Generally, the utility rate of IMV ranged from 18.2% to 38.9% respectively, raising with the monthly percent change (MPC): 1.5% [95% confidence interval (CI): 0.8%, 2.2%] from January 2018 to February 2020 by Joinpoint regression analysis. A continuous decline with the MPC: -1.9% (95% CI: -3.2%, -0.5%) of VAP incidence was demonstrated. However, this trend varied among the different ICUs. We found no significant difference neither in 30-day case fatality nor pathogens of VAP patients. CONCLUSIONS: By Joinpoint regression analysis, we can see February 2020 was an important time point. The surveillance indicators were changed, which influenced the VAP incidence.

Computational modelling of the graft-tunnel interaction in single-bundle ACL reconstructed knee.

OBJECTIVES: Tunnel enlargement and graft failure are common complications associated with ACL reconstruction. The mechanical interaction between the graft and the tunnel aperture may play a more important role. This study aims to evaluate graft position within femoral tunnel and the graft force under external loads. METHODS: An FE model of the femur-graft-tibia complex was constructed from CT images of an anatomically reconstructed knee specimen. The model was subjected to kinematics of passive flexion extension, anterior/posterior translation, internal/external rotation and valgus kinematics, which were collected from experimental testing. Graft shift and rotation of graft-tunnel contact region during flexion/extension and external loadings were recorded and compared to experimental measurements. RESULTS: Model showed that the graft shifted in the femoral tunnel during flexion and under external loads. The graft-tunnel contact area rotated by up to 55° during flexion from full extension to 90° of extension implying that the so-called "wiper effect" occurs during most of flexion angles. CONCLUSIONS: Different regions of the femoral tunnel aperture, particularly the anterior region, were under significantly more contact force from the graft than other areas of the aperture during the anterior translation test, potentially leading to femoral tunnel enlargement to the anterior side of the aperture.

Genome-Wide Analyses of SlFWL Family Genes and Their Expression Profiles under Cold, Heat, Salt and Drought Stress in Tomato.

PLAC8 is a cysteine-rich protein that serves as a central mediator of tumor evolution in mammals. PLAC8 motif-containing proteins widely distribute in fungi, algae, higher plants and animals that have been described to be implicated in fruit size, cell number and the transport of heavy metals such as cadmium or zinc. In tomatoes, FW2.2 is a PLAC8 motif-containing gene that negatively controls fruit size by regulating cell division and expansion in the carpel ovary during fruit development. However, despite FW2.2, other FWL (FW2.2-Like) genes in tomatoes have not been investigated. In this study, we identified the 21 SlFWL genes, including FW2.2, examined their expression profiles under various abiotic adversity-related conditions. The SlFWL gene structures and motif compositions are conserved, indicating that tomato SlFWL genes may have similar roles. Cis-acting element analysis revealed that the SlFWL genes may participate in light and abiotic stress responses, and they also interacted with a variety of phytohormone-responsive proteins and plant development elements. Phylogenetic analyses were performed on five additional plant species, including Arabidopsis, pepper, soybean, rice and maize, these genes were classified into five subfamilies. Based on the results of collinearity analyses, the SlFWL genes have a tighter homologous evolutionary relationship with soybean, and these orthologous FWL gene pairs might have the common ancestor. Expression profiling of SlFWL genes show that they were all responsive to abiotic stresses, each subgroup of genes exhibited a different expression trend. Our findings provide a strong foundation for investigating the function and abiotic stress responses of the SlFWL family genes.

Paraquat-induced ferroptosis suppression via NRF2 expression regulation.

Paraquat (PQ) is an environmentally friendly and efficient herbicide, but PQ misuse or intentional self-use can cause death through multiple organ damage and can cause acute lung injury. Existing clinical treatments alleviate symptoms but do not significantly improve the mortality rate. Ferroptosis is a type of necrosis that presents in a manner very similar to the cell damage induced by high doses of PQ, but the role of ferroptosis in paraquat-induced lung injury remains unclear. In this study, we aimed to explore the role of ferroptosis in PQ-induced A549 cell injury and identify the potential mechanisms and critical sites of protection against PQ-induced A549 injury by ferroptosis inhibitors. We found that the ferroptosis inhibitors Ferr-1 and Lip-1 inhibit ferroptosis by attenuating oxidative stress through the upregulation of NRF2 gene expression. The protective role of the ferroptosis inhibitor Dfo was most evident in paraquat-induced cell injury. Dfo inhibited ferroptosis by iron chelation and promoted NRF2 protein level reduction. NRF2 attenuated PQ-induced ferroptosis in A549 cells, mainly through the upregulation of SLC40A1 to encourage the movement of iron to the extracellular side to alleviate iron overload, and the upregulation of SLC7A11 to promote the expression of GPX4 to inhibit lipid peroxidation.

Diurnal gene oscillations modulated by RNA metabolism in tomato.

Diurnal rhythms are known to regulate the expression of a large number of genes, coordinating plant growth and development with diel changes in light and temperature. However, the impact of RNA metabolism on rhythmic gene oscillations in plant is not yet fully understood. To address this question, we performed transcriptome and degradome profiling on tomato leaves at 6 time points during one 24 h cycle, using RNA-seq and genome-wide mapping of uncapped and cleavage transcripts (GMUCT). Time-series profiling of RNA-seq revealed 9342 diurnal-oscillated genes, which were enriched in various metabolic processes. To quantify the general level of RNA degradation for each gene, we utilized the Proportion Uncapped (PU) metric, which represents the GMUCT/RNA-seq ratio. Oscillated PU analysis revealed that 3885 genes were regulated by rhythmic RNA degradation. The RNA decay of these diurnal genes was highly coordinated with mRNA downregulation during oscillation, highlighting the critical role of internal transcription-degradation balance in rhythmic gene oscillation. Furthermore, we identified 2190 genes undergoing co-translational RNA decay (CTRD) with 5' phosphate read ends enriched at the boundary of ribosomes stalling at translational termination sites. Interestingly, diurnal-changed mRNAs with large amplitudes tended to be co-translationally decay, suggesting that CTRD contributed to the rapid turnover of diurnal mRNAs. Finally, we also identified several genes, whose miRNA cleavage efficiency oscillated in a diurnal manner. Taken together, these findings uncovered the vital functions of RNA metabolism, including rhythmic RNA degradation, CTRD, and miRNA cleavage, in modulating the diurnal mRNA oscillations during diel change at post-transcriptional level in tomato.

Construction and Validation of a Novel Web-Based Nomogram for Solitary Plasmacytoma of Bone of the Spine: A Real-World Analysis Based on the Surveillance, Epidemiology, and End Results Database.

OBJECTIVE: Solitary plasmacytoma of bone of the spine (SPBS) was rarely detected in the past. However, its incidence has gradually increased with improvements in the diagnosis and understanding of the disease. We aimed to conduct a population-based cohort study to characterize the prevalence and factors associated with SPBS and develop a prognostic nomogram for predicting the overall survival of SPBS patients with a real-world analysis based on the surveillance, epidemiology, and end results database. METHODS: Patients with SPBS at diagnosis were identified using the SEER database from 2000-2018. Multivariable and univariate logistic regression analyses were used to identify factors for developing a novel nomogram. Nomogram performance was evaluated using the calibration curve, area under the curve (AUC), and decision curve analyses. Kaplan-Meier analysis was used to estimate survival durations. RESULTS: A total of 1,147 patients were selected for survival analysis. Multivariate analysis demonstrated that independent predictors for SPBS were as follows: ages: 61-74 and 75-94, marital status: unmarried, treatment: radiation alone and radiation with surgery. The 1-, 3-, and 5-year AUCs for OS were 0.733, 0.735, and 0.735 in the training cohort and 0.754, 0.777, and 0.791 in the validation cohort, respectively. The C-index values in the 2 cohorts were 0.704 and 0.729. The results indicated that nomograms could satisfactorily identify patients with SPBS. CONCLUSIONS: Our model effectively demonstrated the clinicopathological features of SPBS patients. The results indicated that the nomogram had a favorable discriminatory ability, good consistency, and yielded clinical benefits for SPBS patients.

Differences in Physiological Responses of Two Tomato Genotypes to Combined Waterlogging and Cadmium Stresses.

Waterlogging and heavy mental (e.g., cadmium) stress are two primary threats to crop growth. The combination of abiotic stresses was common and frequent, especially in the field condition. Even though the effects of individual waterlogging and cadmium on tomato plants have been widely investigated, the response of tomatoes under combined waterlogging and cadmium stress remains unclear. This study aimed to clarify and compare physiological, biochemical characteristics and plant growth of two tomato genotypes under individual and combined stress. Two tomato genotypes ('MIX-002' and 'LA4440') were treated under control, waterlogging, cadmium stress and their combination. The results showed that chloroplast ultrastructure of tomatoes under individual and combined stress was damaged with disordered stroma and grana lamellae. The H2O2 (hydrogen peroxide) content and O2·- (superoxide anion radical) production rate of plants under all the three stresses was not significantly higher than the control except for 'LA4440' under the combined stress. Antioxidant enzymes actively responded in the two tomato genotypes, as shown by significant increase in SOD activity from 'MIX-002' under waterlogging and combined stress and from 'LA4440' under cadmium. Meanwhile, CAT activity of 'MIX-002' under waterlogging and 'LA4440' under combined stress significantly decreased, and the POD activity of 'MIX-002' under combined stress significantly increased as compared with the respective control. The APX activity of 'MIX-002' and 'LA4440' under combined stress was significantly lower and higher than the respective controls. This indicated that tomato plants were able to secure redox homeostasis and protect plants from oxidative damage through the synergetic regulation of antioxidant enzymes. Plant height and biomass of the two genotypes under individual and combined stress significantly decreased, which could be a direct result from the chloroplast alteration and resource re-allocation. Overall, the effects of combined waterlogging and cadmium stress were not simply the sum of individual effects on two tomato genotypes. Distinct ROS (reactive oxygen species) scavenging systems of two tomato genotypes under stresses suggest a genotype-dependent antioxidant enzymes regulation.

Open Versus Arthroscopic Latarjet for Recurrent Anterior Shoulder Instability: A Systematic Review and Meta-analysis.

Background: The open Latarjet (OL) procedure and arthroscopic Latarjet (AL) procedure are able to treat recurrent anterior shoulder instability (RASI) with high success rates. Purpose: To evaluate the clinical efficacy and postoperative revisions and complications between the OL and AL procedures in the treatment of RASI. Study Design: Systematic review; Level of evidence, 3. Methods: MEDLINE, Embase, and the Cochrane Library were searched to retrieve and include cohort studies comparing the OL and AL procedures for RASI. Clinical outcomes were compared, and results were reported as odds ratios (ORs) or mean differences (MDs) with 95% CIs. Results: Eleven clinical trials with 1217 patients were included. There were no differences between the procedures in pain score, Rowe score, Walch-Duplay score, external rotation, persistent apprehension, instability, recurrence, revisions attributed to recurrent instability, overall complications, wound infection, hematoma, graft complications, screw-related complications, or osteoarthritis. When compared with the OL procedure, the AL procedure had a significantly lower nonunion rate (OR, 9.92; 95% CI, 1.71 to 57.71; P = .01); however, the AL procedure had a longer operation time (MD, -24.49; 95% CI, -48.44 to -0.54; P = .05), lower Western Ontario Shoulder Instability Index score (MD, 97.27; 95% CI, 21.91 to 172.63; P = .01), higher revision rate (OR, 0.39; 95% CI, 0.16 to 0.95; P = .04), and greater screw deviation (MD, -6.41; 95% CI, -10.25 to -2.57; P = .001). Conclusion: For most outcome measures, no difference was seen between the OL and AL procedures. The AL procedure had a lower Western Ontario Shoulder Instability Index score and a higher revision rate and appeared to have a significant learning curve. However, the AL procedure resulted in a lower nonunion rate.

miR­424­5p is downregulated in the placentas of patients with preeclampsia and affects trophoblast migration and invasion.

Insufficient invasion of trophoblast cells has been reported to be closely associated with the pathogenesis of preeclampsia (PE). MicroRNAs (miRs) have essential roles in the trophoblasts invasion via targeting specific genes with diverse functions. However, the underlying mechanism remains largely unclear and requires further investigation. The present study aimed to identify and evaluate the potential functions of miRs in trophoblasts invasion and to reveal the underlying mechanisms. In the present study, differentially expressed miRs that were screened based on previously published microarray data (GSE96985) and a significantly downregulated miR-424-5p (miR-424) was chosen for further investigation. Subsequently, reverse transcription-quantitative PCR, CCK-8, apoptosis, wound healing and Transwell assays were performed to determine the cell viability, apoptotic rate, cell migration and invasion of trophoblast cells. The results showed that miR-424 was decreased in placenta specimens from patients with PE. Upregulation of miR-424 promoted cell viability, suppressed cell apoptosis and improved the invasion and migration of trophoblasts, whereas inhibition of miR-424 had opposite results. Adenomatous polyposis coli (APC), a key mediator of Wnt/ß-catenin signaling pathway, was identified as a functional target of miR-424 and an inverse relationship was observed between APC and miR-424 in placenta specimens. Further investigations revealed that APC overexpression efficiently suppressed the effect of miR-424 in trophoblast cells. In addition, the miR-424-mediated effects on trophoblast cells were dependent on the promotion of Wnt/ß-catenin signaling pathway. The present findings revealed that miR-424 regulates the trophoblast cell invasion by regulating Wnt/ß-catenin pathway through targeting APC, indicating miR-424 as a potential candidate for the treatment of PE.

Risk factors for in-hospital mortality and secondary bacterial pneumonia among hospitalized adult patients with community-acquired influenza: a large retrospective cohort study.

BACKGROUND: Secondary bacterial pneumonia is an important complication of seasonal influenza, but little data is available about impact on death and risk factors. This study identified risk factors for all-cause in-hospital mortality and secondary bacterial pneumonia among hospitalized adult patients with community-acquired influenza. METHODS: A retrospective cohort study was performed at a tertiary teaching hospital in southwest China. The study cohort included all adult hospitalized patients with a laboratory-confirmed, community-acquired influenza virus infection during three consecutive influenza seasons from 2017 to 2020. Cause-specific Cox regression was used to analyze risk factors for mortality and secondary bacterial pneumonia, respectively, accounting for competing events (discharge alive and discharge alive or death without secondary bacterial pneumonia, respectively). RESULTS: Among 174 patients enrolled in this study, 14.4% developed secondary bacterial pneumonia and 11.5% died during hospitalization. For all-cause in-hospital mortality, time-varying secondary bacterial pneumonia was a direct risk factor of death (cause-specific hazard ratio [csHR] 3.38, 95% confidence interval [CI] 1.25-9.17); underlying disease indirectly increased death risk through decreasing the hazard of being discharged alive (csHR 0.55, 95% CI 0.39-0.77). For secondary bacterial pneumonia, the final model only confirmed direct risk factors: age ≥ 65 years (csHR 2.90, 95% CI 1.27-6.62), male gender (csHR 3.78, 95% CI 1.12-12.84) and mechanical ventilation on admission (csHR 2.96, 95% CI 1.32-6.64). CONCLUSIONS: Secondary bacterial pneumonia was a major risk factor for in-hospital mortality among adult hospitalized patients with community-acquired influenza. Prevention strategies for secondary bacterial pneumonia should target elderly male patients and critically ill patients under mechanical ventilation.

Effects of nonantibiotic growth promoter combinations on growth performance, nutrient utilization, digestive enzymes, intestinal morphology, and cecal microflora of broilers.

Given the ban on antibiotic growth promoters, the effects of nonantibiotic alternative growth promoter combinations (NAGPCs) on the growth performance, nutrient utilization, digestive enzyme activity, intestinal morphology, and cecal microflora of broilers were evaluated. All birds were fed pellets of two basal diets-starter (0-21 d) and grower (22-42 d)-with either enramycin (ENR) or NAGPC supplemented. 1) control + ENR; 2) control diet (CON, basal diet); 3) control + mannose oligosaccharide (MOS) + mannanase (MAN) + sodium butyrate (SB) (MMS); 4) control + MOS + MAN + Bacillus subtilis (BS) (MMB); 5) control + MOS + fruit oligosaccharide (FOS) + SB (MFS); 6) control + FOS + BS (MFB); 7) control + MOS + FOS + MAN (MFM); 8) control + MOS + BS + phytase (PT) (MBP). ENR, MOS, FOS, SB, MAN, PT, and BS were added at 100, 2,000, 9,000, 1,500, 300, 37, and 500 mg/kg, respectively. The experiment used a completely random block design with six replicates per group: 2400 Ross 308 broilers in the starter phase and 768 in the grower phase. All NAGPCs significantly improved body weight gain (P < 0.01), utilization of dry matter, organic matter, and crude protein (P < 0.05), villus height and villus height/crypt depth in the jejunum and ileum (P < 0.01), and decreased the feed conversion ratio (P < 0.01) at d 21 and 42. MMS, MMB, MFB, and MFM duodenum trypsin, lipase, and amylase activities increased significantly (P < 0.05) at d 21 and 42. On d 21 and 42, MMS, MMB, and MBP increased the abundance of Firmicutes and Bacteroides whereas MMB, MFB, and MBP decreased the abundance of Proteobacteria, compared to ENR and CON. Overall, the NAGPCs were found to have some beneficial effects and may be used as effective antibiotic replacements in broilers.

Comparative Study of Graft Healing in 2 Years after "Tension Suspension" Remnant-Preserving and Non-Remnant-Preserving Anatomical Reconstruction for Sherman Type II Anterior Cruciate Ligament Injury.

PURPOSE: To evaluate the degree of graft healing after "tension suspension" reconstruction of "Sherman II" anterior cruciate ligament injuries versus non-remnant preserving anatomical reconstruction and to compare the clinical outcomes of the two procedures. METHOD: The clinical data of 64 patients were retrospectively included. There were 31 cases in the "tension suspension" remnant-preserving reconstruction group and 33 cases in the non-remnant-preserving anatomical reconstruction group. The International Knee Documentation Committee (IKDC) score, the Tegner score, and the Lysholm activity score were assessed preoperatively and at 6 months, 1 year, and 2 years postoperatively, respectively. The signal/noise quotient (SNQ) of the grafts was measured at 6 months, 1 year, and 2 years after surgery to quantitatively evaluate the maturity of the grafts after ACL reconstruction; the fractional anisotropy (FA) and apparent diffusion coefficient (ADC) of the reconstructed ACL region of interest (ROI) were measured using DTI. RESULT: A total of 64 patients were included in the study. The mean SNQ values of the grafts in the 6 months, 1 year, and 2 years postoperative remnant-preserving reconstruction (RP) groups were lower than those in the non-remnant-preserving (NRP) reconstruction group, with a statistically significant difference (p < 0.05). At each postoperative follow-up, the SNQ values of the tibial and femoral sides of the RP group were lower than those of the NRP group; the SNQ values of the femoral side of the grafts in both groups were higher than those of the tibial side, and the differences were statistically significant (p < 0.05). At 6 months, 1 year, and 2 years postoperatively, the FA and ADC values of the grafts were lower in the RP group than in the NRP group, and the differences were statistically significant (p < 0.05); the IKDC score and Lysholm score of the RP group were higher than the NRP group, which was statistically significant (p < 0.05). CONCLUSION: For Sherman II ACL injury, the graft healing including ligamentization and revascularization at 2 years after the "tension suspension" remnant-preserving reconstruction was better than that of non-remnant-preserving anatomic reconstruction.

miR-124 Exacerbates depressive-like behavior by targeting Ezh2 to induce autophagy.

On the basis of our previous research, miR-124 and autophagy have been shown to be associated with depression and antidepressant treatment, respectively. However, whether miR-124 is involved in depressive-like behavior and antidepressant efficacy through regulating autophagy remains poorly understood. The chronic unpredictable mild stress (CUMS) depression model in mice was established, and then intraperitoneal fluoxetine injections (10 mg/kg) were administered for a duration of 4 weeks. The behavioral changes induced by CUMS were evaluated by the tail suspension test, open field test, sucrose preference test, and elevated plus maze test. Quantitative real-time PCR was used to detect expression levels of miR-124 and its three precursor genes in hippocampus of mice. Western blotting was used to detect the expressions of Ezh2 and autophagy proteins (P62, Atg3, Atg7, LC3-I, and LC3- II) in hippocampus of mice. Depression-like behaviors were successfully induced in CUMS models and reversed by SSRI treatments. The expression levels of miR-124 and its precursor gene ( miR-124-3 ) were significantly increased in the hippocampus of CUMS mice, while the expression levels were significantly decreased after 4 weeks of fluoxetine treatment. The mRNA and protein expressions of Ezh2, a validated target of miR-124, were decreased in the hippocampus of CUMS mice, and the fluoxetine treatment could reverse the expressions. A correlation analysis suggested that miR-124 had a significant negative correlation with Ezh2 mRNA expression. The protein levels of LC3-II/I, P62, and Atg7, which were found to be regulated by Ezh2, were increased in the hippocampus of CUMS mice and decreased after fluoxetine treatment. We speculated that autophagy was enhanced in the CUMS model of depression and might be mediated by miR-124 targeting Ezh2.

Deciphering postnatal limb development at single-cell resolution.

The early postnatal limb developmental progression bridges embryonic and mature stages and mirrors the pathological remodeling of articular cartilage. However, compared with multitudinous research on embryonic limb development, the early postnatal stage seems relatively unnoticed. Here, a systematic work to portray the postnatal limb developmental landscape was carried out by characterization of 19,952 single cells from murine hindlimbs at 4 postnatal stages using single-cell RNA sequencing technique. By delineation of cell heterogeneity, the candidate progenitor sub-clusters marked by Cd34 and Ly6e were discovered in articular cartilage and enthesis, and three cellular developmental branches marked by Col10a1, Spp1, and Tnni2 were reflected in growth plate. The representative transcriptomes and developmental patterns were intensively explored, and the key regulation mechanisms as well as evolvement in osteoarthritis were discussed. Above all, these results expand horizons of postnatal limb developmental biology and reach the interconnections between limb development, remodeling, and regeneration.