Three-dimensional morphological characterization of blood droplets during the dynamic coagulation process.

In this study, we employed a method integrating optical coherence tomography (OCT) with the U-Net and Visual Geometry Group (VGG)-Net frameworks within a convolutional neural network for quantitative characterization of the three dimensional whole blood during the dynamic coagulation process. VGG-Net architecture for the identification of blood droplets across three distinct coagulation stages including drop, gelation, and coagulation achieves an accuracy of up to 99%. In addition, the U-Net architecture demonstrated proficiency in effectively segmenting uncoagulated and coagulated portions of whole blood, as well as the background. Notably, parameters such as volume of uncoagulated and coagulated segments of the whole blood were successfully employed for the precise quantification of the coagulation process, which indicates well for the potential of future clinical diagnostics and analyses.

Exploring the immunological mechanism of Houttuynia cordata in the treatment of colorectal cancer through combined network pharmacology and experimental validation.

To explore the potential mechanisms of Houttuynia cordata in the treatment of CRC using network pharmacology combined with experimental validation. The major active components of Houttuynia cordata were identified using the TCMSP database, and their related targets were mined. CRC-related target genes were obtained through the Genecards and OMIM databases. The R software Ven Diagram package was used for visualization of the intersection of drug and disease targets. The intersection target genes were subjected to GO function enrichment and KEGG pathway enrichment analysis using the R software clusterProfiler package. A "Drug - active component - target - Disease" network was constructed and analyzed using Cytoscape software. Intersection target genes were uploaded to the STRING database, and the resultant data were imported into Cytoscape software to construct a PPI network and filter core target genes. Expression analysis, diagnostic efficacy, and survival analysis were used to demonstrate the function and clinical value of the core target genes. The correlation between core genes in CRC samples and immune cell infiltration was analyzed using the R software and ssGSVA algorithm. Molecular docking validation of core active components with core target genes was performed using AutodockVina 1.2.2 software. Finally, the effects of quercetin and kaempferol, core active components of Houttuynia cordata, on the growth of HCT116 cells and the regulation of core target genes were validated through CCK8 assay, flow cytometry, and RT-qPCR. Seven effective active components and 147 component-related targets were selected, along with 3806 CRC-related target genes. GO analysis mainly involved biological processes such as epithelial cell proliferation, with KEGG pathway analysis focusing on pathways including AGE-RAGE signaling. Quercetin and kaempferol were identified as two core components, with IL1B, MMP9, CXCL8, and IL6 as four core target genes. Immune infiltration analysis showed that IL1B, MMP9, CXCL8, and IL6 primarily exert anti-CRC effects by promoting neutrophil activity. Molecular docking results indicated stable binding capacities of quercetin and kaempferol with IL1B, MMP9, CXCL8, and IL6. Experimental validation showed that quercetin and kaempferol could inhibit the viability of HCT116 cells in a dose-dependent manner, promote apoptosis, and downregulate the expression of IL1B, MMP9, CXCL8, and IL6 genes. Houttuynia cordata may exert therapeutic effects on CRC by modulating the immune microenvironment and anti-inflammatory responses, providing new research directions and theoretical guidance for the treatment of CRC with Houttuynia cordata.

Underlying Mechanism of Lysosomal Membrane Permeabilization in CNS Injury: A Literature Review.

Lysosomes play a crucial role in various intracellular pathways as their final destination. Various stressors, whether mild or severe, can induce lysosomal membrane permeabilization (LMP), resulting in the release of lysosomal enzymes into the cytoplasm. LMP not only plays a pivotal role in various cellular events but also significantly contributes to programmed cell death (PCD). Previous research has demonstrated the participation of LMP in central nervous system (CNS) injuries, including traumatic brain injury (TBI), spinal cord injury (SCI), subarachnoid hemorrhage (SAH), and hypoxic-ischemic encephalopathy (HIE). However, the mechanisms underlying LMP in CNS injuries are poorly understood. The occurrence of LMP leads to the activation of inflammatory pathways, increased levels of oxidative stress, and PCD. Herein, we present a comprehensive overview of the latest findings regarding LMP and highlight its functions in cellular events and PCDs (lysosome-dependent cell death, apoptosis, pyroptosis, ferroptosis, and autophagy). In addition, we consolidate the most recent insights into LMP in CNS injury by summarizing and exploring the latest advances. We also review potential therapeutic strategies that aim to preserve LMP or inhibit the release of enzymes from lysosomes to alleviate the consequences of LMP in CNS injury. A better understanding of the role that LMP plays in CNS injury may facilitate the development of strategic treatment options for CNS injury.

Nirmatrelvir and ritonavir for inpatients with severe or critical COVID-19 beyond five days of symptom onset: a propensity score-matched, multicenter, retrospective cohort study.

BACKGROUND: There is an urgent need for therapeutic strategies for inpatients with severe or critical COVID-19. The evaluation of the clinical benefits of nirmatrelvir and ritonavir (Nmr/r) for these patients beyond five days of symptom onset is insufficient. METHODS: A new propensity score-matched cohort was constructed by using multicenter data from 6695 adult inpatients with COVID-19 from December 2022 to February 2023 in China after the epidemic control measures were lifted across the country. The severity of disease of the inpatients was based on the tenth trial edition of the Guidelines on the Diagnosis and Treatment of COVID-19 in China. The symptom onset of 1870 enrolled severe or critical inpatients was beyond five days, and they received either Nmr/r plus standard treatment or only standard care. The ratio of patients whose SOFA score improved more than 2 points, crucial respiratory endpoints, changes in inflammatory markers, safety on the seventh day following the initiation of Nmr/r treatment, and length of hospital stay were evaluated. RESULTS: In the Nmr/r group, on Day 7, the number of patients with an improvement in SOFA score ≥ 2 was much greater than that in the standard treatment group (P = 0.024) without a significant decrease in glomerular filtration rate (P = 0.815). Additionally, the rate of new intubation was lower (P = 0.004) and the no intubation days were higher (P = 0.003) in the first 7 days in the Nmr/r group. Other clinical benefits were limited. CONCLUSIONS: Our study may provide new insight that inpatients with severe or critical COVID-19 beyond five days of symptom onset benefit from Nmr/r. Future studies, particularly randomized controlled trials, are necessary to verify the above findings.

Syntaxin5 is essential for survival by ensuring midgut epithelial homeostsis and regulating feeding in Locusta migratoria.

Syntaxin5 (Syx5) belongs to SNAREs family, which play important roles in fusion of vesicles to target membranes. Most of what we know about functions of Syx5 originates from studies in fungal or vertebrate cells, how Syx5 operates during the development of insects is poorly understood. In this study, we investigated the role of LmSyx5 in the gut development of the hemimetabolous insect Locusta migratoria. LmSyx5 was expressed in many tissues, with higher levels in the gut. Knockdown of LmSyx5 by RNA interference (RNAi) considerably suppressed feeding in both nymphs and adults. The dsLmSyx5-injected locusts lost body weight and finally died at a mortality of 100%. Furthermore, hematoxylin-eosin staining indicated that the midgut is deformed in dsLmSyx5-treated nymphs and the brush border in midgut epithelial cells is severely damaged, suggesting that LmSyx5 is involved in morphogenesis of the midgut. TEM further showed that the endoplasmic reticulum of midgut cells have a bloated appearance. Taken together, these results suggest that LmSyx5 is essential for midgut epithelial homeostsis that affects growth and development of L. migratoria. Thus, Syx5 is a promising RNAi target for controlling L. migratoria, and even other pests.

Bispecific CAR-T cells targeting FAP and GPC3 have the potential to treat hepatocellular carcinoma.

Chimeric antigen receptor (CAR) T cell therapy has demonstrated robust efficacy against hematological malignancies, but there are still some challenges regarding treating solid tumors, including tumor heterogeneity, antigen escape, and an immunosuppressive microenvironment. Here, we found that SNU398, a hepatocellular carcinoma (HCC) cell line, exhibited high expression levels of fibroblast activation protein (FAP) and Glypican 3 (GPC3), which were negatively correlated with patient prognosis. The HepG2 HCC cell line highly expressed GPC3, while the SNU387 cell line exhibited high expression of FAP. Thus, we developed bispecific CAR-T cells to simultaneously target FAP and GPC3 to address tumor heterogeneity in HCC. The anti-FAP-GPC3 bispecific CAR-T cells could recognize and be activated by FAP or GPC3 expressed by tumor cells. Compared with anti-FAP CAR-T cells or anti-GPC3 CAR-T cells, bispecific CAR-T cells achieved more robust activity against tumor cells expressing FAP and GPC3 in vitro. The anti-FAP-GPC3 bispecific CAR-T cells also exhibited superior antitumor efficacy and significantly prolonged the survival of mice compared with single-target CAR-T cells in vivo. Overall, the use of anti-FAP-GPC3 bispecific CAR-T cells is a promising treatment approach to reduce tumor recurrence caused by tumor antigen heterogeneity.

Effect of thymalfasin on myeloid-derived suppressor cells in patients with non-small cell lung cancer.

OBJECTIVE: To observe the effect of thymalfasin on myeloid-derived suppressor cells (MDSCs) subsets in peripheral blood of patients with non-small cell lung cancer (NSCLC). METHODS: 50 cases of NSCLC (NSCLC group) diagnosed in Chest Hospital of Jiangxi Province were selected as the research subjects, and 50 healthy subjects who underwent physical examination in our hospital during the same period were selected as the healthy control group. The expression of HLA-DR-CD14-CD33+ MDSCs in peripheral blood mononuclear cells and tumor tissue single cell suspension of NSCLC patients before and after thymalfasin treatment was explored by flow cytometry. RESULTS: The proportion of MDSCs in peripheral blood of NSCLC group was 1.70±0.52%, which was significantly higher than that in peripheral blood (0.51±0.15%) of healthy controls (P < 0.05). The proportion of HLA-DR-CD14-CD33+ MDSCs in the tissues of NSCLC group was 1.65±0.43% before treatment and 1.15±0.50% after treatment (P < 0.05). The proportion of MDSCs in peripheral blood of NSCLC patients before treatment was 1.70±0.52%, and that after treatment was 0.59±0.18% (P < 0.05). CONCLUSION: Thymalfasin can reduce the number of MDSCs in peripheral blood mononuclear cells. The application of thymalfasin in the treatment of NSCLC patients can help to enhance the anti-tumor effect.

Exploratory metabolomic analysis for characterizing the metabolic profile of the urinary bladder under estrogen deprivation.

Background: Estrogen homeostasis is crucial for bladder function, and estrogen deprivation resulting from menopause, ovariectomy or ovarian dysfunction may lead to various bladder dysfunctions. However, the specific mechanisms are not fully understood. Methods: We simulated estrogen deprivation using a rat ovariectomy model and supplemented estrogen through subcutaneous injections. The metabolic characteristics of bladder tissue were analyzed using non-targeted metabolomics, followed by bioinformatics analysis to preliminarily reveal the association between estrogen deprivation and bladder function. Results: We successfully established a rat model with estrogen deprivation and, through multivariate analysis and validation, identified several promising biomarkers represented by 3, 5-tetradecadiencarnitine, lysoPC (15:0), and cortisol. Furthermore, we explored estrogen deprivation-related metabolic changes in the bladder primarily characterized by amino acid metabolism imbalance. Conclusion: This study, for the first time, depicts the metabolic landscape of bladder resulting from estrogen deprivation, providing an important experimental basis for future research on bladder dysfunctions caused by menopause.

Sodium alginate/gelatin hydrogel spheres loaded with Fructus Ligustri Lucidi essential oil: Preparation, characterization and biological activity.

The application of plant essential oils in the food industry is often hindered by their poor water solubility and high volatilize. Encapsulation has emerged as an effective solution to this problem. This study focuses on the preparation of Fructus Ligustri Lucidi essential oil gel spheres (FEOH) based sodium alginate and gelatin. The optimum formulation for FEOH was established by Box-Behnken Design response surface testing, resulting in a composition of 10 % FEO, 5 % TW20 and 2 % CaCl2. This formulation achieved an encapsulation efficiency of 85.56 %. FTIR and SEM results indicated the successful encapsulation of FEO within the gel spheres. Furthermore, DSC and TGA results showed that encapsulation enhanced the thermal stability of the essential oil. At room temperature, the water content of FEOH exceeded 90 %, and it showed the highest swelling ratio of 62.5 % in an alkaline medium at different pH conditions. The in vitro release behavior showed that FEOH was released up to 85.28 % in oil-based food simulants within 2 h. FEOH showed strong antibacterial activity, with a Minimum Inhibitory Concentration (MIC) of 128 mg/mL against Staphylococcus aureus and 256 mg/mL against Escherichia coli. The gel spheres obtained in this research show significant potential as food preservatives in food matrices.

Effects of perineal massage at different stages on perineal and postpartum pelvic floor function in primiparous women: a systematic review and meta-analysis.

BACKGROUND: Perineal massage, as a preventive intervention, has been shown to reduce the risk of perineal injuries and may have a positive impact on pelvic floor function in the early postpartum period. However, there is still debate concerning the best period to apply perineal massage, which is either antenatal or in the second stage of labor, as well as its safety and effectiveness. Meta-analysis was used to evaluate the effect of implementing perineal massage in antenatal versus the second stage of labor on the prevention of perineal injuries during labor and early postpartum pelvic floor function in primiparous women. METHODS: We searched nine different electronic databases from inception to April 16, 2024. The randomized controlled trials (RCTs) we included assessed the effects of antenatal and second-stage labor perineal massage in primiparous women. All data were analyzed with Revman 5.3, Stata Statistical Software, and Risk of Bias 2 was used to assess the risk of bias. Subgroup analyses were performed based on the different periods of perineal massage. The primary outcomes were the incidence of perineal integrity and perineal injury. Secondary outcomes were perineal pain, duration of the second stage of labor, postpartum hemorrhage, urinary incontinence, fecal incontinence, and flatus incontinence. RESULTS: This review comprised a total of 10 studies that covered 1057 primigravid women. The results of the analysis showed that perineal massage during the second stage of labor reduced the perineal pain of primigravid women in the immediate postpartum period compared to the antenatal period, with a statistical value of (MD = -2.29, 95% CI [-2.53, -2.05], P < 0.001). Additionally, only the antenatal stage reported that perineal massage reduced fecal incontinence (P = 0.04) and flatus incontinence (P = 0.01) in primiparous women at three months postpartum, but had no significant effect on urinary incontinence in primiparous women at three months postpartum (P = 0.80). CONCLUSIONS: Reducing perineal injuries in primiparous women can be achieved by providing perineal massage both antenatally and during the second stage of labor. Pelvic floor function is improved in the postnatal phase by perineal massage during the antenatal stage. TRIAL REGISTRATION: CRD42023415996 (PROSPERO).

Precisely targeted drug delivery by mesenchymal stem cells-based biomimetic liposomes to cerebral ischemia-reperfusion injured hemisphere.

The precise and targeted delivery of therapeutic agents to the lesion sites remains a major challenge in treating brain diseases represented by ischemic stroke. Herein, we modified liposomes with mesenchymal stem cells (MSC) membrane to construct biomimetic liposomes, termed MSCsome. MSCsome (115.99 ± 4.03 nm) exhibited concentrated accumulation in the cerebral infarcted hemisphere of mice with cerebral ischemia-reperfusion injury, while showing uniform distribution in the two cerebral hemispheres of normal mice. Moreover, MSCsome exhibited high colocalization with damaged nerve cells in the infarcted hemisphere, highlighting its advantageous precise targeting capabilities over liposomes at both the tissue and cellular levels. Leveraging its superior targeting properties, MSCsome effectively delivered Dl-3-n-butylphthalide (NBP) to the injured hemisphere, making a single-dose (15 mg/kg) intravenous injection of NBP-encapsulated MSCsome facilitate the recovery of motor functions in model mice by improving the damaged microenvironment and suppressing neuroinflammation. This study underscores that the modification of the MSC membrane notably enhances the capacity of liposomes for precisely targeting the injured hemisphere, which is particularly crucial in treating cerebral ischemia-reperfusion injury.

Objectively Measured Physical Activity In Children With Developmental Coordination Disorder: A Systematic Review And Meta-analysis.

OBJECTIVE: To conduct a systematic review and meta-analysis to understand the difference in objectively measured physical activities (PA) between children with and without developmental coordination disorder (DCD). DATA SOURCES: A systematic literature search from four databases (PubMed, Science Direct, Web of Science, and Cochrane library) was conducted in July 2023. STUDY SELECTION: Studies that met the following criteria were considered: (1) the studies should classified children with DCD based on DSM-IV, DSM-IV-TR, or DSM-V diagnosis criteria, (2) the studies aimed to evaluate PA using objective measurements and provided the amount of time spent in PA and/or SB, (3) a control group of TD children was recruited, (4) the full-text article was written in English. DATA EXTRACTION: The following data from all included studies were extracted: the first author's surname and published year, study design, country, total sample size, the measure of PA, the intensity of PA, categories of PA level and main finding(s). DATA SYNTHESIS: 12 articles met the inclusion criteria for the systematic review, 10 of which were further entered into the meta-analysis. Overall mean difference in moderate to vigorous PA (MVPA) between two groups was -0.17 (95% CI: -0.25 to -0.09, I2 = 48.7%, p = 0.029). When subgroup analysis of age was further conducted (i.e., school-aged vs. preschool), a significant pooled effect size with no heterogeneity was found in school-aged children (i.e., 6-14 years old) (standardized mean difference (SMD) = -0.27, 95% CI: -0.38 to -0.16, I2 = 43.1%, p = 0.08). CONCLUSIONS: Children with DCD spent significantly less time participating in MVPA, specifically those children aging between 6 and 14 years. These findings help raise the awareness for the parents and physicians toward insufficient participation in PA in children with DCD.

Influence of biodegradable plastics on the generation of disinfection byproducts in the chlorination process.

Biodegradable plastics (BPs) have seen a continuous increase in annual production and application due to their environmentally sustainable characteristics. However, research on the formation of disinfection byproducts (DBPs) from biodegradable microplastics (BMPs) during chlorination is limited, and the effects of aqueous solution chemistry on this process have yet to be explored. Therefore, two biodegradable microplastics, polylactic acid (PLA) and polybutylene adipate terephthalate (PBAT), were investigated in this study to examine the changes in their physicochemical properties before and after chlorination, and the formation of DBPs under different environmental conditions. The results showed that PLA was more chlorine-responsive, and generated more DBPs. The pH converted some of the intermediates into more stable DBPs by affecting the concentration of HClO and base-catalyzed reactions, whereas ionic strength slightly reduced DBP concentration by ion adsorption and promoting the aggregation of BMPs. Finally, since PLA has a slightly greater volume of mesopores and micropores compared to PBAT, it may more effectively adsorb DBP precursors beyond natural organic matter (NOM), such as some anthropogenic pollutants, thus potentially decreasing the formation of chlorinated DBPs in surface water. This research explored the potentiality for DBP formation by BMPs under different water quality conditions during the disinfection process, which is useful for assessing the environmental hazards of BMPs.

Ligand-Accelerated Pd-Catalyzed C(sp2)-H and C(sp3)-H Arylation under the Catalytic System.

It is universally acknowledged that ligands can improve the reaction activity to simplify the reaction operating conditions and enrich the applicability of the reaction. Therefore, we developed N-octylglycine ligand-accelerated Pd-catalyzed ortho-arylation of benzoic acids under mild conditions with just 6 h; moreover, this N-octylglycine ligand was successfully implemented to carboxyl-directed Pd-catalyzed ß-C(sp3)-H arylation and ortho-arylation of phenylacetic acids under mild conditions.

A radiomics-based nomogram may be useful for predicting telomerase reverse transcriptase promoter mutation status in adult glioblastoma.

BACKGROUND AND PURPOSE: As a crucial diagnostic and prognostic biomarker, telomerase reverse transcriptase (TERT) promoter mutation holds immense significance for personalized treatment of patients with glioblastoma (GBM). In this study, we developed a radiomics nomogram to determine the TERT promoter mutation status and assessed its prognostic efficacy in GBM patients. METHODS: The study retrospectively included 145 GBM patients. A comprehensive set of 3736 radiomics features was extracted from preoperative magnetic resonance imaging, including T2-weighted imaging, T1-weighted imaging (T1WI), contrast-enhanced T1WI, and fluid-attenuated inversion recovery. The construction of the radiomics model was based on integrating the radiomics signature (rad-score)with clinical features. Receiver-operating characteristic curve analysis was employed to evaluate the discriminative ability of the prediction model, and the risk score was used to stratify patient outcomes. RESULTS: The least absolute shrinkage and selection operator classifier identified 10 robust features for constructing the prediction model, and the radiomics nomogram exhibited excellent performance in predicting TERT promoter mutation status, with area under the curve values of.906 (95% confidence interval [CI]:.850-.963) and.899 (95% CI:.708-.966) in the training and validation sets, respectively. The clinical utility of the radiomics nomogram is further supported by calibration curve and decision curve analyses. Additionally, the radiomics nomogram effectively stratified GBM patients with significantly different prognoses (HR = 1.767, p = .019). CONCLUSION: The radiomics nomogram holds promise as a modality for evaluating TERT promoter mutations and prognostic outcomes in patients with GBM.

The Prognostic Value of Intravascular Ultrasound-Guided Coronary Endothelial Diameter in Percutaneous Coronary Intervention: The UNIQUE-PCI Outcome Study.

Objectives: In the context of coronary artery disease (CAD) and percutaneous coronary intervention (PCI), it is essential to explore the variations in coronary endothelial diameter. Understanding these variations holds significance for the early diagnosis and treatment of CAD. This study aims to elucidate the relevance of coronary endothelial diameter in the broader landscape of CAD and PCI outcomes. Methods: This study is a retrospective cohort study. Consecutive patients with CAD who underwent PCI during hospitalization were included. IVUS measured the endothelial diameter of the coronary artery, and the patients were divided into the endothelial diameter constricted group, normal group and dilated group. The primary endpoint event was all-cause death, and the secondary endpoint events were cardiac death, non-fatal myocardial infarction, and elective revascularization. Follow-up was completed from August 2021 to February 2022. The follow-up period from August 2021 to February 2022 was selected to capture a comprehensive view of long-term postoperative outcomes in coronary artery disease patients undergoing PCI, ensuring a contemporary and relevant assessment of the study endpoints. Results: The study ultimately included 705 patients, which included 295 (41.8%) in the endothelial diameter constricted group, 410 (58.2%) in the endothelial diameter normal group and 221 (31.3%) in the endothelial diameter dilated group. Patients with dilated endothelial diameter experienced a reduction in 5-year all-cause mortality, cardiac mortality, non-fatal heart attack, and elective revascularization rates. Specifically, the 5-year all-cause mortality rate in the dilated group was 1.79 (95% CI: 1.07-3.00), cardiac mortality was 3.73 (95% CI: 1.27-10.95), non-fatal heart attack rate was 1.65 (95% CI: 0.99-2.75), and elective revascularization rate was 2.15 (95% CI: 1.30-3.60) (P < .05). The Cox proportional-hazards model indicated that age, AMI, and endothelial diameter expansion were identified as risk factors for 5-year all-cause mortality (P < .05). There was no statistically significant difference in the 5-year all-cause mortality, cardiogenic mortality, elective revascularization rate and non-fatal myocardial infarction rate. Conclusions: Patients with dilated endothelial diameter had decreased 5-year all-cause mortality, cardiac mortality, non-fatal heart attack, and elective revascularization rates. IVUS evaluation of the diameter of the coronary endothelium prior to PCI can aid in the classification and prevention of CAD risk. The retrospective design and potential biases associated with hospitalization data, along with the absence of certain clinical parameters, should be considered when interpreting the findings. The IVUS evaluation of coronary endothelial diameter not only aids in risk classification but also has the potential to inform personalized treatment strategies, enhancing patient care in coronary artery disease.

Key node identification for a network topology using hierarchical comprehensive importance coefficients.

Key nodes are similar to important hubs in a network structure, which can directly determine the robustness and stability of the network. By effectively identifying and protecting these critical nodes, the robustness of the network can be improved, making it more resistant to external interference and attacks. There are various topology analysis methods for a given network, but key node identification methods often focus on either local attributes or global attributes. Designing an algorithm that combines both attributes can improve the accuracy of key node identification. In this paper, the constraint coefficient of a weakly connected network is calculated based on the Salton indicator, and a hierarchical tenacity global coefficient is obtained by an improved K-Shell decomposition method. Then, a hierarchical comprehensive key node identification algorithm is proposed which can comprehensively indicate the local and global attributes of the network nodes. Experimental results on real network datasets show that the proposed algorithm outperforms the other classic algorithms in terms of connectivity, average remaining edges, sensitivity and monotonicity.

Genetic Screening Revealed the Negative Regulation of miR-310~313 Cluster Members on Imd Pathway during Gram-Negative Bacterial Infection in Drosophila.

Innate immune response is the first line of host defense against pathogenic microorganisms, and its excessive or insufficient activation is detrimental to the organism. Many individual microRNAs (miRNAs) have emerged as crucial post-transcriptional regulators of immune homeostasis in Drosophila melanogaster. However, the synergistical regulation of miRNAs located within a cluster on the Imd-immune pathway remains obscured. In our study, a genetic screening with 52 transgenic UAS-miRNAs was performed to identify ten miRNAs or miRNA clusters, including the miR310~313 cluster, which may function on Imd-dependent immune responses. The miRNA RT-qPCR analysis showed that the expression of miR-310~313 cluster members exhibited an increase at 6-12 h post E. coli infection. Furthermore, the overexpression of the miR-310~313 cluster impaired the Drosophila survival. And the overexpression of miR-310/311/312 reduced Dpt expression, an indication of Imd pathway induced by Gram-negative bacteria. Conversely, the knockdown of miR-310/311/312 led to increases in Dpt expression. The Luciferase reporter expression assays and RT-qPCR analysis confirmed that miR-310~313 cluster members directly co-targeted and inhibited Imd transcription. These findings reveal that the members of the miR-310~313 cluster synergistically inhibit Imd-dependent immune responses by co-targeting the Imd gene in Drosophila.

Toward Designing Reactive Metal Clusters for Dinitrogen Activation: A Guideline Based on N2 Initial Adsorption.

Gas-phase metal clusters are ideal models to explore transition-metal-mediated N2 activation mechanism. However, the effective design and search of reactive clusters in N2 activation are currently hindered by the lack of clear guidelines. Inspired by the Sabatier principle, we discovered in this work that N2 initial adsorption energy (ΔEads) is an important parameter to control the N2 activation reactivity of metal clusters in the gas phase. This mechanistic insight obtained from high-level calculations rationalizes the N2 activation reactivity of many previously reported metal clusters when combined with the known factor determining the N≡N cleavage process. Furthermore, based on this guideline of ΔEads, we successfully designed several new reactive clusters for cleaving N≡N triple bond under mild conditions, including FeV2S2-, TaV2C2-, and TaV2C3-, the high N2 activation reactivity of which has been fully corroborated in our gas phase experiments employing mass spectrometry with collision-induced dissociation. The importance of ΔEads revealed in this work not only reshapes our understanding of N2 activation reactions in the gas phase but also could have implication for other N2 activation processes in the condensed phase. The more general establishment of this new perspective on N2 activation reactivity warrants future experimental and computational studies.