CLPs-miR-103a-2-5p inhibits proliferation and promotes cell apoptosis in AML cells by targeting LILRB3 and Nrf2/HO-1 axis, regulating CD8 + T cell response.

BACKGROUND: LILRB3, a member of the leukocyte immunoglobulin-like receptor B (LILRB) family, has immunosuppressive functions and directly regulates cancer development, which indicates that LILRB3 is an attractive target for cancer diagnosis and therapy. Novel therapeutic treatments for acute myeloid leukemia (AML) are urgent and important, and RNA therapeutics including microRNAs (miRNAs) could be an effective option. Here, we investigate the role of dysregulated miRNA targeting LILRB3 in the AML microenvironment. METHODS: Potential miRNAs binding to the 3'-untranslated region (3'-UTR) of the LILRB3 mRNA were predicted by bioinformatics websites. Then, we screened miRNAs targeting LILRB3 by quantitative real-time PCR, and the dual luciferase reporter assay. The expression of LILRB3 and microRNA (miR)-103a-2-5p in AML were determined and then their interactions were also analyzed. In vitro, the effects of miR-103a-2-5p were determined by CCK8, colony formation assay, and transwell assay, while cell apoptosis and cell cycle were analyzed by flow cytometry. Cationic liposomes (CLPs) were used for the delivery of miR-103a-2-5p in the AML mouse model, which was to validate the potential roles of miR-103a-2-5p in vivo. RESULTS: LILRB3 was upregulated in AML cells while miR-103a-2-5p was dramatically downregulated. Thus, a negative correlation was found between them. MiR-103a-2-5p directly targeted LILRB3 in AML cells. Overexpressed miR-103a-2-5p significantly suppressed the mRNA and protein levels of LILRB3, thereby inhibiting AML cell growth and reducing CD8 + T cell apoptosis. In addition, overexpressed miR-103a-2-5p reduced both the relative expression of Nrf2/HO-1 pathway-related proteins and the ratio of GSH/ROS, leading to the excessive intracellular ROS that may promote AML cell apoptosis. In the mouse model, the delivery of miR-103a-2-5p through CLPs could inhibit tumor growth. CONCLUSIONS: MiR-103a-2-5p serves as a tumor suppressor that could inhibit AML cell proliferation and promote their apoptosis by downregulating LILRB3 expression, suppressing the Nrf2/HO-1 axis, and reducing the ratio of GSH/ROS. Besides, our findings indicate that miR-103a-2-5p may enhance the CD8 + T cell response by inhibiting LILRB3 expression. Therefore, the delivery of miR-103a-2-5p through CLPs could be useful for the treatment of AML.

Synthesis and Photocatalytic Application of Magnetic CoFe2O4/Conjugated Poly(vinyl chloride) Derivative Nanocomposite.

CoFe2O4 has potential for application as a magnetically recoverable visible-light photocatalyst, but its photocatalytic activity is encumbered by the high recombination probability of its photogenerated holes (h+) and electrons (e-). This work was undertaken to boost the photocatalysis of CoFe2O4 through coupling with conjugated poly(vinyl chloride) derivative (CPVC). An easily implementable solvothermal-liquid solid mixing-evaporation of the solvent-pyrolysis method was exploited to synthesize CoFe2O4/CPVC nanocomposites. The photocatalytic capabilities of the products were assessed through photocatalyzing the reduction of Cr(VI) under visible-light (λ > 420 nm). The results demonstrate that the optimal CoFe2O4/CPVC nanocomposite (CoFe2O4/CPVC-2) has markedly heightened photocatalytic activity (3.6 times that of CoFe2O4) and competent reusability and is magnetically recoverable. Furthermore, CoFe2O4/CPVC-2 also shows superior performance toward photocatalytic treatment of the diluted Cr(VI)-containing passivation solution of copper alloys. It is deduced based on the photoelectricity measurement results that the increased photocatalysis of CoFe2O4/CPVC-2 is chiefly attributed to its p-n heterojunction structure, which greatly elevates the h+-e- separation and transfer efficiency. When waste PVC plastic films (replacing the new pure PVC powder) were utilized for the synthesis, the obtained CoFe2O4/CPVC nanocomposite exhibited even better photocatalytic activity (4 times that of CoFe2O4). This work not only has made a new magnetically recoverable, efficient visible-light photocatalyst for decontamination of Cr(VI) in water but also is inspirational for recycling PVC plastic waste to produce high-valued visible-light photocatalysts.

The Association of Preoperative Diabetes With Postoperative Delirium in Older Patients Undergoing Major Orthopedic Surgery: A Prospective Matched Cohort Study.

BACKGROUND: Postoperative delirium (POD) is a common form of postoperative brain dysfunction, especially in the elderly. However, its risk factors remain largely to be determined. This study aimed to investigate whether (1) preoperative diabetes is associated with POD after elective orthopedic surgery and (2) intraoperative frontal alpha power is a mediator of the association between preoperative diabetes and POD. METHODS: This was a prospective matched cohort study of patients aged 60 years or more, with a preoperative diabetes who underwent elective orthopedic surgery. Nondiabetic patients were matched 1:1 to diabetic patients in terms of age, sex, and type of surgery. Primary outcome was occurrence of POD, assessed using the 3-minute Diagnostic Confusion Assessment Method (3D-CAM) once daily from 6 pm to 8 pm during the postoperative days 1-7 or until discharge. Secondary outcome was the severity of POD which was assessed for all participants using the short form of the CAM-Severity. Frontal electroencephalogram (EEG) was recorded starting before induction of anesthesia and lasting until discharge from the operating room. Intraoperative alpha power was calculated using multitaper spectral analyses. Mediation analysis was used to estimate the proportion of the association between preoperative diabetes and POD that could be explained by intraoperative alpha power. RESULTS: A total of 138 pairs of eligible patients successfully matched 1:1. After enrollment, 6 patients in the diabetes group and 4 patients in the nondiabetes group were excluded due to unavailability of raw EEG data. The final analysis included 132 participants with preoperative diabetes and 134 participants without preoperative diabetes, with a median age of 68 years and 72.6% of patients were female. The incidence of POD was 16.7% (22/132) in patients with preoperative diabetes vs 6.0% (8/134) in patients without preoperative diabetes. Preoperative diabetes was associated with increased odds of POD after adjustment of age, sex, body mass index, education level, hypertension, arrhythmia, coronary heart disease, and history of stroke (odds ratio, 3.2; 95% confidence interval [CI], 1.4-8.0; P = .009). The intraoperative alpha power accounted for an estimated 20% (95% CI, 2.6-60%; P = .021) of the association between diabetes and POD. CONCLUSIONS: This study suggests that preoperative diabetes is associated with an increased risk of POD in older patients undergoing major orthopedic surgery, and that low intraoperative alpha power partially mediates such association.

Exploring ways to improve China's ecological well-being amidst air pollution challenges using mixed methods.

Ecological well-being performance (EWP), a novel concept in sustainable development research, diverges from traditional ecological efficiency in terms of perspectives, core content, and driving factors. However, research on EWP remains insufficiently comprehensive, particularly the corresponding theoretical and methodological investigations into driving pathways. To address this gap, this study develops an "economy-environment-health" framework, incorporating air pollution and associated health losses into the evaluation system, and employs a two-stage Super-NSBM and Window DEA model for reevaluating EWP. The study further investigates the primary pathways of EWP, focusing on environmental regulations, technological innovation, and structural adjustments through both quantitative and qualitative methods. Quantitative spatial econometric analysis reveals that factors such as market-driven environmental regulations, green invention patents, and industrial and energy consumption structures significantly enhance EWP. While examining the "net effects" contributions of individual variables using spatial econometric models, the fsQCA method is employed to identify four effective driving paths for EWP from a configurational perspective. These paths are 1) technological innovation and structural adjustment under environmental regulations with public participation; 2) a combination of environmental regulation, technological innovation, and structural adjustment; 3) structural adjustment with minimal influence from environmental regulations and technological innovation; and 4) structural adjustment directed by market-incentive environmental regulations.

Water-Immiscible Coacervate as a Liquid Magnetic Robot for Intravascular Navigation.

Developing magnetic ultrasoft robots to navigate through extraordinarily narrow and confined spaces like capillaries in vivo requires synthesizing materials with excessive deformability, responsive actuation, and rapid adaptability, which are difficult to achieve with the current soft polymeric materials, such as elastomers and hydrogels. We report a magnetically actuatable and water-immiscible (MAWI) coacervate based on the assembled magnetic core-shell nanoparticles to function as a liquid robot. The degradable and biocompatible millimeter-sized MAWI coacervate liquid robot can remain stable under changing pH and salt concentrations, release loaded cargoes on demand, squeeze through an artificial capillary network within seconds, and realize intravascular targeting in vivo guided by an external magnetic field. We believe the proposed "coacervate-based liquid robot" can implement demanding tasks beyond the capability of conventional elastomer or hydrogel-based soft robots in the field of biomedicine and represents a distinct design strategy for high-performance ultrasoft robots.

KRAS mutations upregulate Runx1 to promote occurrence of head and neck squamous cell carcinoma.

Gene mutations play an important role in head and neck squamous cell carcinoma (HNSCC) by not only promoting the occurrence and progression of HNSCC but also affecting sensitivity to treatment and prognosis. KRAS is one of the most frequently mutated oncogenes, which has been reported to have a mutation rate from 1.7% to 12.7% and may lead to poor prognosis in HNSCC, but its role remains unclear. Here, we found that the KRAS mutation can promote HNSCC generation through synergism with 4-Nitroquinoline-1-Oxide(4NQO). Mechanistically, KRAS mutations can significantly upregulate Runx1 to promote oral epithelial cell proliferation and migration and inhibit apoptosis. Runx1 inhibitor Ro 5-3335 can effectively inhibit KRAS-mutated HNSCC progression both in vitro and in vivo. These findings suggest that the KRAS mutation plays an important role in HNSCC and that Runx1 may be a novel therapeutic target for KRAS-mutated HNSCC.

A Polymer Acceptor with Grafted Small Molecule Acceptor Unit for Efficient All Polymer Organic Solar Cells.

A polymer acceptor, named PX-1, is  designed and synthesized using a polymerization strategy with grafted small molecule acceptors. This design approach allows for the freedom of end groups while maintaining efficient terminal packing, enhancing π-π interactions, and facilitating charge transport. All-polymer organic solar cells based on PM6: PX-1 demonstrate a promising efficiency of 13.55%. The result presents an alternative pathway for the design of high-efficiency polymer acceptors through the careful regulation of small molecule acceptor monomers and linker units.

Structural basis of phosphorylation-induced activation of the response regulator VbrR.

Two-component systems typically consist of a paired histidine kinase and response regulator and couple environmental changes to adaptive responses. The response regulator VbrR from Vibrio parahaemolyticus, a member of the OmpR/PhoB family, regulates virulence and antibiotic resistance genes. The activation mechanism of VbrR remains unclear. Here, we report the crystal structures of full-length VbrR in complex with DNA in the active conformation and the N-terminal receiver domain (RD) and the C-terminal DNA-binding domain (DBD) in both active and inactive conformations. Structural and biochemical analyses suggest that unphosphorylated VbrR adopts mainly as inactive dimers through the DBD at the autoinhibitory state. The RD undergoes a monomer-to-dimer transition upon phosphorylation, which further induces the transition of DBD from an autoinhibitory dimer to an active dimer and enables its binding with target DNA. Our study suggests a new model for phosphorylation-induced activation of response regulators and sheds light on the pathogenesis of V. parahaemolyticus.

Noninvasive Real-Time Monitoring of Wetting Progression in Membrane Distillation Using Impedance Spectroscopy.

Membrane distillation (MD) is a promising technology for the treatment of high salinity wastewater using a hydrophobic membrane; however, the occurrence of wetting due to surfactants in polluted or low surface tension liquid impedes MD application. Common monitoring approaches, such as conductivity and flux measurement, cannot explain the wetting phenomenon that occurs during the wetting process in detail. Recently, impedance spectroscopy has been proposed for early wetting detection, as it depends on the change of water/air composition in the membrane pores. An earlier and larger variation was observed with precise signal detection. In this study, we proposed an analytical approach to estimate the wetting front, which is the average feed intrusion distance, by the impedance value recorded in real-time operation. With this proposed approach, the wetting mechanism in the presence of a surfactant and the effect of pore size on a commercial polyvinylidene fluoride membrane could be quantified, which cannot be explained in detail using conductivity and flux measurements.

Health-related quality of life in osteoarthritis patients: a systematic review and meta-analysis.

The aim of this meta-analysis was to compare health-related quality of life (HRQoL) of Osteoarthritis (OA) patients and controls. A systematic literature search was performed on PubMed, Web of Science and EMBASE from database inception to 7 January 2020. Random effect model was performed to summarize the scores of each domain and the forest plot was used to compare the scores of OA patients with healthy controls. Subgroup analyses were conducted to explore the source of heterogeneity. Statistical analyses were executed using Review Manager (version 5.1). In total, six studies were included in this study, including 7094 patients with OA and 12 100 healthy controls, which were all reliable to summarize the scores of the Medical Outcomes Study 36-item Short-Form Health Survey (SF-36). Meta-analyses found that pooled mean HRQoL score for the SF-36 each domain (physical function, physical role function, body pain, general health, vitality, social function, emotional role function, mental health) was lower in patients with OA than in healthy controls, especially the score in the dimension of physical role function. OA have a substantial impact HRQoL. HRQoL is a significant component of measuring overall health, which contributes to formulate successful self-disease management plan, patient-centered care, and develop effective interventions target confidence.

Effects of Mn2+ and humic acid on microbial community structures, functional genes for nitrogen and phosphorus removal, and heavy metal resistance genes in wastewater treatment.

Considering the wide occurrence of Mn2+ and humic acid (HA) in environmental media, the effects of Mn2+ (5-16 mg/L) and HA (10 mg/L) on microbial community structures, functional genes for nitrogen and phosphorus removal, and heavy metal resistance genes (HMRGs) were investigated in wastewater treatment using sequencing batch bioreactors (SBRs). The treatment efficiencies of influent chemical oxygen demands (COD), NH4+-N, and PO43--P were unaffected during the entire operational processes irrespective of whether Mn2+ and HA were supplied. Although the functional prediction of genetic information via sequencing analysis showed that the microbial activity was not influenced by Mn2+ and HA from different SBRs, the abundance of dominant phyla (Proteobacteria, Actinobacteriota, Firmicutes, and Bacteroidota), classes (Saccharimonadia, Gammaproteobacteria, and Bacilli), and genera (unidentified_Chloroplast, TM7a, Micropruina, Candidatus_Competibacter, Lactobacillus, OLB12, and Pediococcus) was different. Compared to the SBR without Mn2+ and HA supplementation, the abundance of functional genes for nitrogen and phosphorus removal (narG, nirS, nosZ, ppk, and phoD) and HMRGs (corA and mntA) significantly increased under Mn2+ stress, but significantly decreased with the addition of HA except for genes nirS and ppk. The abundance of genes corA and mntA was related to the partially dominant microbes and functional genes, and might be reduced by supplying HA. This study provides insight into the effects of Mn2+ and HA on functional genes for nitrogen and phosphorus removal and HMRGs in wastewater treatment.

A Phenanthrocarbazole-Based Dopant-Free Hole-Transport Polymer with Noncovalent Conformational Locking for Efficient Perovskite Solar Cells.

Adequate hole mobility is the prerequisite for dopant-free polymeric hole-transport materials (HTMs). Constraining the configurational variation of polymer chains to afford a rigid and planar backbone can reduce unfavorable reorganization energy and improve hole mobility. Herein, a noncovalent conformational locking via S-O secondary interaction is exploited in a phenanthrocarbazole (PC) based polymeric HTM, PC6, to fix the molecular geometry and significantly reduce reorganization energy. Systematic studies on structurally explicit repeats to targeted polymers reveals that the broad and planar backbone of PC remarkably enhances π-π stacking of adjacent polymers, facilitating intermolecular charge transfer greatly. The inserted "Lewis soft" oxygen atoms passivate the trap sites efficiently at the perovskite/HTM interface and further suppress interfacial recombination. Consequently, a PSC employing PC6 as a dopant-free HTM offers an excellent power conversion efficiency of 22.2 % and significantly improved longevity, rendering it as one of the best PSCs based on dopant-free HTMs.

BioContainers Registry: Searching Bioinformatics and Proteomics Tools, Packages, and Containers.

BioContainers is an open-source project that aims to create, store, and distribute bioinformatics software containers and packages. The BioContainers community has developed a set of guidelines to standardize software containers including the metadata, versions, licenses, and software dependencies. BioContainers supports multiple packaging and container technologies such as Conda, Docker, and Singularity. The BioContainers provide over 9000 bioinformatics tools, including more than 200 proteomics and mass spectrometry tools. Here we introduce the BioContainers Registry and Restful API to make containerized bioinformatics tools more findable, accessible, interoperable, and reusable (FAIR). The BioContainers Registry provides a fast and convenient way to find and retrieve bioinformatics tool packages and containers. By doing so, it will increase the use of bioinformatics packages and containers while promoting replicability and reproducibility in research.

Saline and alkaline stresses alter soil properties and composition and structure of gene-based nitrifier and denitrifier communities in a calcareous desert soil.

BACKGROUND: Saline and alkaline stresses damages the health of soil systems. Meanwhile, little is known about how saline or alkaline stress affects soil nitrifier and denitrifier communities. Therefore, we compared the responses of gene-based nitrifier and denitrifier communities to chloride (CS), sulfate (SS), and alkaline (AS) stresses with those in a no-stress control (CK) in pots with a calcareous desert soil. RESULTS: Compared with CK, saline and alkaline stress decreased potential nitrification rate (PNR) and NO3-N; increased pH, salinity, water content, and NH4-N; and decreased copy numbers of amoA-AOA and amoA-AOB genes but increased those of denitrifier nirS and nosZ genes. Copies of nirK increased in SS and AS but decreased in CS. There were more copies of amoA-AOB than of amoA-AOA and of nirS than of nirK or nosZ. Compared with CK, SS and AS decreased operational taxonomic units (OTUs) of amoA-AOB but increased those of nirS and nosZ, whereas CS decreased nirK OTUs but increased those of nosZ. The numbers of OTUs and amoA-AOB genes were greater than those of amoA-AOA. There were positive linear relations between PNR and amoA-AOA and amoA-AOB copies. Compared with CK, the Chao 1 index of amoA-AOA and amoA-AOB decreased in AS, that of nirK increased in CS and SS, but that of nirS and nosZ increased in all treatments. The Shannon index of amoA-AOB decreased but that of nirS increased in CS and SS, whereas the index of nirK decreased in all treatments. Saline and alkaline stress greatly affected the structure of nitrifier and denitrifier communities and decreased potential biomarkers of nirS-type; however, AS increased those of nirK- and nosZ-type, and SS decreased those of nosZ-type. Soil water content, pH, and salinity were important in shaping amoA-AOA and denitrifier communities, whereas soil water and pH were important to amoA-AOB communities. CONCLUSION: These results indicate that the nitrifier and denitrifier communities respond to saline and alkaline stresses conditions. Communities of amoA-AOA and amoA-AOB contribute to nitrification in alluvial gray desert soil, and those of nirS are more important in denitrification than those of nirK or nosZ.

Psychological status and disease activity were major predictors of body image disturbances in patients with Sjögren's syndrome.

Sjögren's syndrome (SS) often leads to disease-related body defects and functional impairments, which may result in the body image disturbances (BID) of patients. The aim of this study was to investigate the severity and predictors of BID among SS patients. Two hundred and thirty-one SS patients [mean (IQR) age: 51 (42-58); females: 94.4%] and 224 age and sex-matched healthy controls were included in this cross-sectional study. Questionnaires were applied: body image disturbance questionnaire (BIDQ), the hospital anxiety and depression scale (HADS), fatigue severity scale (FSS), the 10-cm pain Visual Analog Scale (VAS), the oral health impact profile-14 (OHIP-14), the ocular surface disease index (OSDI), the social support rate scale (SSRS). Independent sample t-test, Mann-Whitney U-test, Chi-square test, spearman rank correlation, and stepwise linear regression were performed by SPSS version 20.0 to analyze these data. In 231 SS patients, the mean of the overall BIDQ score was 1.80 ± 1.21, and SS patients had significantly higher scores in each domain of BIDQ compared with healthy controls (p < 0.05). The stepwise multiple linear regression analysis revealed that high BIDQ score was predicted by severe anxiety (ß = 0.081; p < 0.001), high disease activity (ß = 0.038; p < 0.001) and poor oral health (ß = 0.017; p = 0.007) in SS patients. Patients with SS suffer from severe BID and it is necessary for rheumatologists to pay more attention to SS patients' body image disturbance, especially those with high disease activity, severe anxiety, and poor oral health to improve patients' quality of life.

Serum urate goal attainment and associated factors in Chinese gout patients.

We conducted a cross-sectional study with 350 gout patients to investigate serum urate acid (sUA) goal attainment (sUA < 360 µmol/L) and associated factors in Chinese gout patients in the Affiliated Hospital of Nantong University from August 2015 to September 2017. Descriptive statistics, health assessment questionnaire, global visual analog scale for general health and Gout Knowledge Questionnaire were calculated comparing patients at sUA goal or not at sUA goal. Univariate analysis and logistic regression models were applied to analyze data. The proportion of urate-lowering therapy (ULT) use was 61.2% (211/350). The mean ± standard deviation sUA of the participants was 475.07 ± 121.53 µmol/L. Only 17.4% of gout patients attain the serum urate goal, which also means 289(82.6%) patients cannot keep their sUA below the target of 360 µmol/L. The factors associated with sUA goal attainment including age, education level and ULT use. Among these, age, waist hip ratio, ULT, family history of cardiovascular disease was the predictor of sUA goal attainment. In conclusion, more than 80% of gout patients did not achieve sUA goal. Younger and fatter patients were more difficult to control the serum urate level.

Gout and risk of diabetes mellitus: meta-analysis of observational studies.

To determine the prevalence of diabetes mellitus (DM) in people with gout, and investigate the relationship between gout and the occurrence of DM. Systematic review and meta-analysis of epidemiological studies. Data sources: MEDLINE, Web of Science, EMBASE and CINAHL databases, hand-searched reference lists, citation history and contact with authors. Eligibility criteria: cohort, case-control or cross-sectional studies which examined the occurrence of DM amongst adults with gout (with or without gout group) in primary care or general population samples. Prevalence and risk estimate meta-analyses were performed using a random-effects model. A total of 23 identified studies matched the inclusion criteria, reporting on a total of 575 284 gout patients. Meta-analyses revealed that the prevalence of DM in gout patients was 16% (95% CI, 14-18%, I2 = 99.8%) according to clinical interviews. In the subgroup analysis, the prevalence of DM was higher in the female population (18%, 95%CI 2.7-33.3%) than the male population (12.6%, 95%CI 8.2-17.1%). As age increased, the incidence of diabetes in gout population increased. DM is commonly found among patients with gout. Patients with gout should be actively screened for DM and its consequences.

Electrospun Nanofiber Membranes Incorporating PDMS-Aerogel Superhydrophobic Coating with Enhanced Flux and Improved Antiwettability in Membrane Distillation.

Electrospun nanofiber membranes (ENMs) have garnered increasing interest due to their controllable nanofiber structure and high void volume fraction properties in membrane distillation (MD). However, MD technology still faces limitations mainly due to low permeate flux and membrane wetting for feeds containing low surface tension compounds. Perfluorinated superhydrophobic membranes could be an alternative, but it has negative environmental impacts. Therefore, other low surface energy materials such as silica aerogel and polydimethylsiloxane (PDMS) have great relevancy in ENMs fabrication. Herein, we have reported the high flux and nonwettability of ENMs fabricated by electrospraying aerogel/polydimethylsiloxane (PDMS)/polyvinylidene fluoride (PVDF) over electrospinning polyvinylidene fluoride- co-hexafluoropropylene (PVDF-HFP) membrane (E-PH). Among various concentrations of aerogel, the 30% aerogel (E-M3-A30) dual layer membrane achieved highest superhydrophobicity (∼170° water contact angle), liquid entry pressure (LEP) of 129.5 ± 3.4 kPa, short water droplet bouncing performance (11.6 ms), low surface energy (4.18 ± 0.27 mN m-1) and high surface roughness ( Ra: 5.04 µm) with re-entrant structure. It also demonstrated nonwetting MD performance over a continuous 7 days operation of saline water (3.5% of NaCl), high antiwetting with harsh saline water containing 0.5 mM sodium dodecyl sulfate (SDS, 28.9 mN m-1), synthetic algal organic matter (AOM).

LL-37 inhibits LPS-induced inflammation and stimulates the osteogenic differentiation of BMSCs via P2X7 receptor and MAPK signaling pathway.

Oral diseases, such as periapical periodontitis and periodontitis, are characterized by inflammation-induced bone loss. LL-37, a human antimicrobial peptide (AMP), has multiple biological functions and the potential to promote osteogenesis. Therefore, this study aimed to investigate the regulatory effects of LL-37 within normal and inflammatory microenvironments. The roles of P2X7 receptor (P2X7R) and mitogen-activated protein kinase (MAPK) signaling pathway were also demonstrated. The results showed that LL-37 promoted bone marrow stromal cell (BMSC) proliferation, migration and osteogenic differentiation. LL-37 inhibited the expression of the inflammatory cytokines interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α) and receptor activator of nuclear factor kappa-B ligand (RANKL) at both protein and gene levels, and attenuated the lipopolysaccharide (LPS)-induced inhibition of osteogenesis. Immunofluorescence (IF) confirmed P2X7R expression in BMSCs. BBG, a P2X7R antagonist, significantly attenuated LL-37-promoted osteogenesis. The phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun NH2-terminal kinase (JNK) increased after LL-37 stimulation, which did not affect p38 phosphorylation. The effects of LL-37 on osteogenesis-related gene expression were markedly attenuated by selective inhibitors of ERK1/2 and JNK. Furthermore, a mouse model of LPS-stimulated calvarial osteolysis was established, and results showed that LL-37 markedly inhibited osteoclastic bone resorption. In conclusion, we speculate that LL-37 inhibits inflammation and promotes BMSC osteogenesis via P2X7R and MAPK signaling pathway.

Integrated whole-genome and transcriptome sequence analysis reveals the genetic characteristics of a riboflavin-overproducing Bacillus subtilis.

Commercial riboflavin production with Bacillus subtilis has been developed by combining rational and classical strain development for almost two decades, but how an improved riboflavin producer can be created rationally is still not completely understood. In this study, we demonstrate the combined use of integrated genomic and transcriptomic analysis of the genetic basis for riboflavin over-production in B. subtilis. This methodology succeeded in discerning the positive mutations in the mutagenesis derived riboflavin producer B. subtilis 24/pMX45 through whole-genome sequencing and transcriptome sequencing. These included RibC (G199D), ribD+(G+39A), PurA (P242L), CcpN(A44S), YvrH (R222Q) and two nonsense mutations YhcF (R90*) and YwaA (Q68*). Reintroducing these specific mutations into the wild-type strain recovered the riboflavin overproduction phenotype and subsequent metabolic engineering greatly improved riboflavin production, achieving an up to 3.4-fold increase of the riboflavin titer over the sequenced producer. A novel mutation, YvrH (R222Q), involved in a typical two-component regulatory system deregulated the purine de novo synthesis pathway and increased the pool of intracellular purine metabolites, which in turn increased riboflavin production. Taken together, we present a case study of combining genome and transcriptome analysis to elucidate the genetic underpinnings of a complex cellular property, which enabled the transfer of beneficial mutations to engineer a reference strain into an overproducer.