The metabolic activation of pentachlorophenol to chloranil as a potent inhibitor of human and rat placental 3ß-hydroxysteroid dehydrogenases.

Pentachlorophenol (PCP) is a widely used pesticide. However, whether PCP and its metabolite chloranil have endocrine-disrupting effects by inhibiting placental 3ß-hydroxysteroid dehydrogenase 1 (3ß-HSD1) remains unclear. The study used in vitro assays with human and rat placental microsomes to measure 3ß-HSD activity as well as human JAr cells to evaluate progesterone production. The results showed that PCP exhibited moderate inhibition of human 3ß-HSD1, with an IC50 value of 29.83 µM and displayed mixed inhibition in terms of mode of action. Conversely, chloranil proved to be a potent inhibitor, demonstrating an IC50 value of 147 nM, and displaying a mixed mode of action. PCP significantly decreased progesterone production by JAr cells at 50 µM, while chloranil markedly reduced progesterone production at ≥1 µM. Interestingly, PCP and chloranil moderately inhibited rat placental homolog 3ß-HSD4, with IC50 values of 27.94 and 23.42 µM, respectively. Dithiothreitol (DTT) alone significantly increased human 3ß-HSD1 activity. Chloranil not PCP mediated inhibition of human 3ß-HSD1 activity was completely reversed by DTT and that of rat 3ß-HSD4 was partially reversed by DTT. Docking analysis revealed that both PCP and chloranil can bind to the catalytic domain of 3ß-HSDs. The difference in the amino acid residue Cys83 in human 3ß-HSD1 may explain why chloranil is a potent inhibitor through its interaction with the cysteine residue of human 3ß-HSD1. In conclusion, PCP is metabolically activated to chloranil as a potent inhibitor of human 3ß-HSD1.

Mechanical Stretch-Induced ATP Release from Osteocytes Promotes Osteogenesis of Bone Marrow Mesenchymal Stem Cells.

BACKGROUND: Mandibular distraction osteogenesis (MDO) is a highly effective method for bone regeneration, commonly employed in treating craniofacial defects and deformities. Osteocytes sense mechanical forces in the pericellular space, relay external stimuli to biochemical changes, and send signals to other effector cells, including bone marrow mesenchymal stem cells (BM-MSCs), to regulate bone resorption and formation. Piezo1 potentially affects the secretion signal molecules of bone cells under mechanical stretch. The primary aim of this study was to enhance our comprehension of the molecular biology underlying this therapeutic approach and to identify specific signaling molecules that facilitate bone formation in response to stretch forces. METHODS: Mechanical stretching was applied to negative controls and Piezo1 knockdown osteocyte-like MLO-Y4 cells. Alkaline phosphatase and Alizarin Red S staining were used to survey the osteogenic potential of BM-MSCs. The production and secretion content of adenosine triphosphate (ATP) was measured using ATP content determination analysis. Pathway-related and osteo-specific genes and proteins were evaluated using real-time polymerase chain reaction (RT-PCR), Western blots, and immunofluorescence. Mitochondrial organization was examined with a transmission electron microscope. RESULTS: The conditioned medium of stretch-exposed MLO-Y4s significantly upregulated osteogenesis-related indicators of BM-MSCs (p < 0.001). The upregulation of BM-MSC osteogenesis was associated with ATP release from osteocytes. Mechanically induced calcium transfer and transcriptional coactivator with PDZ-binding motif (TAZ) nuclear translocation mediated by Piezo1 could promote mitochondrial fission and ATP release. Osteocytes detected stretch forces through Piezo1, triggering calcium influx, TAZ nuclear translocation, and ATP production. CONCLUSIONS: The stretch stimulation of Piezo1 induces calcium influx, which in turn promotes calcium-related TAZ nuclear translocation, changes in mitochondrial dynamics, and the release of ATP in osteocytes. This signaling cascade leads to an up-regulation in the osteogenic capacity of BM-MSCs. Mitochondrial energy metabolism of mechanosensitive protein Piezo1-dependent and ATP release may provide a new effective intervention method for mechanically related bone remodeling.

S100A4 modulates cell proliferation, apoptosis and fibrosis in the hyperplastic prostate.

Benign prostatic hyperplasia (BPH) is one of the most common diseases in elderly men worldwide that may result in lower urinary tract symptoms (LUTS). At present, the specific pathophysiological mechanism for BPH/LUTS LUTS remains unclear. S100 calcium binding protein A4 (S100A4), a member of the calcium binding protein family, regulates a variety of biological processes including cell proliferation, apoptosis and fibrosis. The aim of the current study was to explore and clarify the possible role of S100A4 in BPH/LUTS. The human prostate stromal cell line (WPMY-1), rat prostate epithelial cells, human prostate tissues and two BPH rat models were employed in this study. The expression and localization of S100A4 were detected by quantitative real time PCR (qRT-PCR), immunofluorescence microscopy, Western blotting and immunohistochemistry analysis. Also, S100A4 knockdown or overexpression cell models were constructed and a BPH rat model was induced with testosterone propionate (T) or phenylephrine (PE). The BPH animals were treated with Niclosamide, a S100A4 transcription inhibitor. Results demonstrated that S100A4 was mainly localized in human prostatic stroma and rat prostatic epithelium, and showed a higher expression in BPH. Knockdown of S100A4 induced cell apoptosis, cell proliferation arrest and a reduction of tissue fibrosis markers. Overexpression of S100A4 reversed the aforementioned changes. We also demonstrated that S100A4 regulated proliferation and apoptosis mainly through the ERK pathway and modulated fibrosis via Wnt/ß-catenin signaling. In conclusion, our novel data demonstrate that S100A4 could play a crucial role in BPH development and may be explored as a new therapeutic target of BPH.

A novel lysosome-related prognostic signature associated with prognosis and immune infiltration landscape in oral squamous cell carcinoma.

Background: Predicting the outcome of oral squamous cell carcinoma (OSCC) is challenging due to its diverse nature and intricate causes. This research explores how lysosome-associated genes (LRGs) might forecast overall survival (OS) and correlate with immune infiltration in OSCC patients. Methods: We analyzed OSCC patients' LRGs' mRNA expression data and clinical details from The Cancer Genome Atlas (TCGA) and Gene Expression Omnibus (GEO). Through univariate Cox regression, we pinpointed LRGs with prognostic potential. A signature comprising 12 LRGs linked to prognosis was developed via the Least Absolute Shrinkage and Selection Operator (LASSO) in a training dataset. Patients were classified as higher or lower risk based on their risk scores, and the prognostic independence of the risk score was assessed using multivariate analysis. The model's robustness and precision were confirmed through bioinformatics in the GEO test set. Differential gene expression analysis between risk groups highlighted functional disparities, while various immune evaluation methods elucidated immune differences. Results: The prognostic framework utilized 12 LRGs (SLC46A3, MANBA, NEU1, SDCBP, BRI3, TMEM175, CD164, GPC1, SFTPB, TPP1, Biglycan (BGN) and TMEM192), showing that higher risk was associated with poorer OS. This set of genes independently predicted OS in OSCC, linking LRGs to cellular adhesion and extracellular matrix involvement. Initial assessments using ssGSEA and CIBERSORT suggested that the adverse outcomes in the higher-risk cohort may be tied to immune system deregulation. Conclusion: Twelve-LRGs signature has been identified for OSCC prognosis prediction, offering novel directions for lysosome-targeted therapies against OSCC.

Non-pharmacological interventions for preventing suicide attempts: A systematic review and network meta-analysis.

Suicide attempts can cause serious physical harm or death. It would be crucial to gain a better understanding of the comparative efficacy of non-pharmacological interventions. We aimed to identify which non-pharmacological interventions are more effective in preventing suicide attempts. PubMed, Web of Science, and EMBASE databases were searched systematically from their inception until 3 April 2023. To be eligible for inclusion, randomized controlled trials (RCTs) had to meet the following criteria: Participants were individuals who had suicidal ideation or a history of severe self-harm or attempted suicide. A network meta-analysis was performed using a random effects model to estimate the treatment effect of various non-pharmacological interventions. (PROSPERO registration number: CRD42023411393). We obtained data from 54 studies involving 17,630 participants. Our primary analysis found that Cognitive therapy (CT) (OR=0.19, 95%CI =0.04-0.81), Dialectical Behavior Therapy (DBT) (OR=0.37, 95%CI =0.13-0.97), Cognitive-behavioral therapy (CBT) (OR=0.42, 95%CI =0.17-0.99), and Brief intervention and contact (BIC) (OR=0.65, 95%CI=0.44-0.94) were superior to TAU (within the longest available follow-up time) in preventing suicide attempts, while other intervention methods do not show significant advantages over TAU. Secondary analysis showed that the two intervention measures (CT and BIC) were effective when follow-up time did not exceed 6 months, but there was no effective intervention measure with longer follow-up times. CT, DBT, CBT, and BIC have a better effect in preventing suicide attempts than other non-pharmacological interventions. Additional research is necessary to validate which interventions, as well as which combinations of interventions, are the most effective.

Pseudo-targeted lipidomics insights into lipid discrepancies between yak colostrum and mature milk based on UHPLC-Qtrap-MS.

Yak milk is essential to maintain the normal physiological functions of herders in Tibetan areas of China. However, the lipid components of yak colostrum (YC) and mature milk (YM) have not been systematically studied. We employed a quantitative lipidomics to comprehensively describe the alterations in the milk lipid profile of lactating yaks. Herein, totally 851 lipids from 28 lipid subclasses in YC and YM were identified and screened for 43 significantly different lipids (SDLs; variable importance in projection > 1, fold change < 0.5 or > 2 with P < 0.05), with cholesterol ester (CE, 16:0) and triacylglycerol (TAG, 54:6 (20:5), 50:1 (16:0), 56:6 (20:5)) were the potential lipid biomarkers. Fourteen SDLs were modulated downwards, and 29 SDLs were modulated upwards in YM. Moreover, by analyzing lipid metabolic pathways in these SDLs, glycerophospholipid metabolism was the most critical. Our results furnish integral lipid details for evaluating yak milk's nutritional quality.

Screening of Lactiplantibacillus plantarum NML21 and Its Maintenance on Postharvest Quality of Agaricus bisporus through Anti-Browning and Mitigation of Oxidative Damage.

Browning and other undesirable effects on Agaricus bisporus (A. bisporus) during storage seriously affect its commercial value. In this study, a strain, Lactiplantibacillus plantarum NML21, that resists browning and delays the deterioration of A. bisporus was screened among 72 strains of lactic acid bacteria (LAB), and its preservative effect was analyzed. The results demonstrated that gallic acid, catechin, and protocatechuic acid promoted the growth of NML21, and the strain conversion rates of gallic acid and protocatechuic acid reached 97.16% and 95.85%, respectively. During a 15 d storage of the samples, the NML21 treatment displayed a reduction in the browning index (58.4), weight loss (2.64%), respiration rate (325.45 mg kg-1 h-1), and firmness (0.65 N). The treatment further inhibited Pseudomonas spp. growth and polyphenol oxidase activity, improved the antioxidant capacity, reduced the accumulation of reactive oxygen species, and reduced the malonaldehyde content and cell membrane conductivity. Taken together, the optimized concentrations of NML21 may extend the shelf life of A. bisporus for 3-6 d and could be a useful technique for preserving fresh produce.

The 3' Non-Coding Sequence Negatively Regulates PD-L1 Expression, and Its Regulators Are Systematically Identified in Pan-Cancer.

The 3'-untranslated region (3'-UTR) of PD-L1 is significantly longer than the coding sequences (CDSs). However, its role and regulators have been little studied. We deleted whole 3'-UTR region by CRISPR-Cas9. Prognostic analysis was performed using online tools. Immune infiltration analysis was performed using the Timer and Xcell packages. Immunotherapy response prediction and Cox regression was performed using the R software. MicroRNA network analysis was conducted by the Cytoscape software. The level of PD-L1 was significantly and dramatically up-regulated in cells after deleting the 3'-UTR. Additionally, we discovered a panel of 43 RNA-binding proteins (RBPs) whose expression correlates with PD-L1 in the majority of cancer cell lines and tumor tissues. Among these RBPs, PARP14 is widely associated with immune checkpoints, the tumor microenvironment, and immune-infiltrating cells in various cancer types. We also identified 38 microRNAs whose individual expressions are associated with PD-L1 across different cancers. Notably, miR-3139, miR-4761, and miR-15a-5p showed significant associations with PD-L1 in most cancer types. Furthermore, we revealed 21 m6A regulators that strongly correlate with PD-L1. Importantly, by combining the identified RBP and m6A regulators, we established an immune signature consisting of RBMS1, QKI, ZC3HAV1, and RBM38. This signature can be used to predict the responsiveness of cancer patients to immune checkpoint blockade treatment. We demonstrated the critical role of the 3'-UTR in the regulation of PD-L1 and identified a significant number of potential PD-L1 regulators across various types of cancer. The biomarker signature generated from our findings shows promise in predicting patient prognosis. However, further biological investigation is necessary to explore the potential of these PD-L1 regulators.

Preparation, characterization, and in vivo evaluation of glycyrrhetinic acid-mediated nano-drug delivery system co-loaded with syringopicroside and hydroxytyrosol.

This study aimed to create a glycyrrhetinic acid (GA)-mediated, multi-component, self-assembled nano-drug delivery system co-loaded with syringopicroside (S) and hydroxytyrosol (H) obtained from Syringa Linn by synthesizing a GA-polyethylene glycol-poly (lactic acid-co-glycolic acid) (GPP) nanoparticle delivery carrier to actively target the liver. The nanoparticles were optimized using the central composite design. Nanoparticle characterization, cytotoxicity, pharmacodynamics, and tissue distribution study were performed. The optimized SH-GPP nanoparticle was a white solid powder, which was safe and non-toxic. The particle size and Zeta potential of the nanoparticles were 101.5 ± 3.18 nm and -23.3 ± 0.82 mV, respectively. The polydispersity index value (PDI) was 0.190 ± 0.005; the particle size distribution was comparatively uniform. The average total encapsulation efficiency of the optimized SH-GPP nanoparticle was 50.26% ± 1.29%, and drug loading was 15.47% ± 0.39%. After S and H were arranged into nanoparticles, the proliferation inhibition of HepG2.2.15 cells was improved, and the aim of drug-loaded synergism between GPP and SH was achieved. The GA-mediated nanoparticles were better targeted, were retained longer in vivo, and had higher concentrations in the liver than the unmodified nanoparticles. These nanoparticles have the potential to be a new effective anti-hepatitis B treatment and have great research potential in clinical treatment.

Magnetic resin composites for the enrichment of proteins, peptides and phosphopeptides.

There is growing interest in the development of materials for enriching proteins and phosphoproteins from complex sample matrices for mass spectrometric analysis. Herein, we designed and synthesized two types of magnetic resin composites, i.e., MTS9200@Fe3O4 and FPA90CL@Fe3O4, and assessed their applications as adsorbents for enriching proteins, peptides and phosphopeptides. With the combination of Fe3+-IMAC interaction (MTS9200) or electrostatic attraction (FPA90CL) of resins and the adsorption of Fe3O4, the prepared composites exhibited higher capacities for adsorbing a protein (bovine serum albumin, at 195.71 and 135.03 mg g-1 for MTS9200@Fe3O4 and FPA90CL@Fe3O4, respectively) than MTS9200, FPA90CL and Fe3O4. In addition, due to the contributions of the hydrophobic skeleton of resins and Fe3O4, the magnetic resin composites allowed for efficient enrichment of peptides. Moreover, through Fe3+-IMAC interaction or electrostatic attraction of resins and Fe-O MOAC interaction of Fe3O4 with phosphate groups, phosphopeptides could also be captured. Furthermore, we employed the prepared composites for enriching proteins and phosphopeptides from human serum, where 466 and 506 proteins, and 434 and 356 phosphorylation sites, were detected from human serum after being processed with FPA90CL@Fe3O4 and MTS9200@Fe3O4, respectively. Together, our work revealed the great potential of magnetic resin composites as enrichment materials for proteomics and phosphoproteomics analysis.

Observation of a Folded Dirac Cone in Heavily Doped Graphene.

Superlattice potentials imposed on graphene can alter its Dirac states, enabling the realization of various quantum phases. We report the experimental observation of a replica Dirac cone at the Brillouin zone center induced by a superlattice in heavily doped graphene with Gd intercalation using angle-resolved photoemission spectroscopy (ARPES). The replica Dirac cone arises from the (√3× âˆš3)R30° superlattice formed by the intervalley coupling of two nonequivalent valleys (e.g., the Kekulé-like distortion phase), accompanied by a bandgap opening. According to the findings, the replica Dirac band in Gd-intercalated graphene disappears beyond a critical temperature of 30 K and can be suppressed by potassium adsorption. The modulation of the replica Dirac band is primarily attributable to the residual frozen gas, which can act as a source of intervalley scattering at temperatures below 30 K. Our results highlight the persistence of the hidden Kekulé-like phase within the heavily doped graphene, enriching our current understanding of its replica Dirac Fermions.

Analysis of the effect of Tenebrio Molitor rennet on the flavor formation of Cheddar cheese during ripening based on gas chromatography-ion mobility spectrometry (GC-IMS).

This study aimed to evaluate the potential application of Tenebrio Molitor rennet (TMR) in Cheddar cheese production, and to use gas chromatography-ion mobility spectrometry (GC-IMS) to monitor flavor compounds and fingerprints of cheese during ripening. The results indicated that Cheddar cheese prepared from TMR (TF) has fat content significantly lower than that of commercial rennet (CF) (p < 0.05). However, the results of the sensory evaluation showed that there were no statistically significant differences between the two kinds of cheese (p > 0.05). Both cheeses were rich in free amino acids and free fatty acids. Compared to the CF cheese, gamma-aminobutyric acid and Ornithine contents of the TF cheese reached 187 and 749 mg/kg, respectively, during 120 days of ripening. Moreover, GC-IMS provided information on the characteristics of 40 flavor substances (monomers and dimers) in the TF cheese during ripening. Only 30 flavor substances were identified in the CF cheese. The fingerprint of the two kinds of cheese during ripening can be established by GC-IMS and principal component analysis based on the identified flavor compounds. Therefore, TMR has potential application in Cheddar cheese production. GC-IMS might be applied for the quick, accurate and comprehensive monitoring of cheese flavor during ripening.

Analysis of 16 mycotoxins in genuine traditional Chinese medicine for five medicinal parts: Classification of analytical method based on PANI@CS extraction-UPLC-MS/MS.

A novel PANI@CS solid-phase dispersive extractant combined with ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was developed for the first time, which was used for high-throughput, multi-component, real-time online rapid pretreatment and quantitative classification of 16 mycotoxins from five different medicinal parts of 13 genuine traditional Chinese medicines (TCMs). Ultra performance liquid chromatography combined with triple quadrupole mass spectrometry was used for separation and ESI detection. An internal standard isotope matching calibration was used for quantification purposes to compensate for matrix effects. The limits of detection (LOD) of 16 mycotoxins ranged from 0.1 to 6.0 µg/kg. The linear coefficients (R2) were ≥0.996 in the linear range from 10.0 to 200 µg/L. The recoveries of the 16 mycotoxins ranged from 90.1% to 105.8%, and the relative standard deviations (RSDs) ranged from 1.3% to 4.1%. Thirteen TCMs from five representative medicinal parts were selected and tested under the best sample preparation procedure and chromatographic analysis conditions. The results showed that the method could improve the sensitivity and accuracy of the sample analysis, improve the selectivity and reproducibility of the decolorization and purification of TCMs, which is suitable for the practical application of mycotoxin in trace analysis. This method can also provide a new idea for accurate, efficient, rapid and multi-component online detection of mycotoxins for quality and safety control of TCMs.

Sensitive Current Sensor Based on a Lanthanide Framework with Lewis Basic Bipyridyl Sites for Cu2+ Detection.

A new Yb-based three-dimensional metal-organic framework with free Lewis basic sites, [Yb2(ddbpdc)3(CH3OH)2] (referred to as ACBP-6), from YbCl3 and (6R,8R)-6,8-dimethyl-7,8-dihydro-6H-[1,5]dioxonino[7,6-b:8,9-b']dipyridine-3,11-dicarboxylic acid (H2ddbpdc) was synthesized by a conventional solvothermal method. Two Yb3+ are connected by three carboxyl groups to form the [Yb2(CO2)5] binuclear unit, which is further bridged by two carboxyl moieties to produce a tetranuclear secondary building unit. With further ligation of the ligand ddbpdc2-, a 3-D MOF with helical channels is constructed. In the MOF, Yb3+ only coordinates with O atoms, leaving the bipyridyl N atoms of ddbpdc2- unoccupied. The unsaturated Lewis basic sites make this framework possible to coordinate with other metal ions. After growing the ACBP-6 in situ into a glass micropipette, a novel current sensor is formed. This sensor shows high selectivity and a high signal-to-noise ratio toward Cu2+ detection with a detection limit of 1 µM, due to the stronger coordination ability between the Cu2+ and the bipyridyl N atoms.

Semicarbazide-based fluorescent probe for detection of Cu2+ and formaldehyde in different channels.

Two fluorescent sensors with the receptor semicarbazide respectively at 7- (CAA) and 3-position (CAB) of coumarin were designed and synthesized. CAA exhibits fluorescence turn-on response to Cu2+ by triggering the intramolecular charge transfer (ICT) process via Cu2+-catalyzed hydrolysis, and can detect formaldehyde (FA) at different channel by inhibiting the photo-induced electron transfer (PET). However, CAB displays quite different responses: the photophysical properties hardly changed in the presence of FA; while a three-stage fluorescence response of fast quenching, steady increasing and slowly decreasing was found upon addition of Cu2+. The high selectivity enabled CAA a good candidate for quantification of Cu2+ and formaldehyde as well as bioimaging Cu2+ in living cells. Good linear relationships between the fluorescence intensity and analyte concentration were observed in the range of 0.1-30 µM for Cu2+ and 1.0-50 µM for FA, and their detection limits (LOD) were calculated to be 0.43 µM and 1.92 µM (3δ/k), respectively.

Changes of maxillary central incisor and alveolar bone in Class II Division 2 nonextraction treatment with a fixed appliance or clear aligner: A pilot cone-beam computed tomography study.

INTRODUCTION: This retrospective clinical study investigated the clinical changes of maxillary central incisor and alveolar bone in Class II Division 2 nonextraction treatment with fixed appliances or clear aligners on the basis of cone-beam computed tomography. METHODS: Fifty-nine Chinese Han patients with similar demographic characteristics were collected from a conventional bracket group, a self-ligating bracket group, and a clear aligner group. All measurements about root resorption and alveolar bone thickness on the cone-beam computed tomography images were tested. Changes between pretreatment and posttreatment were evaluated by paired-sample t test. The variation among the 3 groups was compared by 1-way analysis of variance. RESULTS: The resistance center of the maxillary central incisor showed upward or forward movement, and the axial inclination was increased in 3 groups (P <0.0001). Root volume loss in the clear aligner group (23.68 ± 4.82 mm3) was significantly less than that in the fixed appliances group (28.24 ± 6.44 mm3 in the conventional bracket group, 28.17 ± 6.07 mm3 in the self-ligating bracket group) (P <0.05). All 3 groups showed a significant decrease in palatal alveolar bone and total bone thickness at all 3 levels at posttreatment. In contrast, labial bone thickness significantly increased except for crestal level l. Among the 3 groups, the clear aligner group had a prominent increase in labial bone thickness at the apical level (P = 0.0235). CONCLUSIONS: Clear aligner treatment for Class II Division 2 malocclusions could effectively reduce the incidence of fenestration and root resorption. Our findings will be beneficial to comprehensively understand the effectiveness of different appliances for Class II Division 2 malocclusions treatment.

Atf4 regulates angiogenic differences between alveolar bone and long bone macrophages by regulating M1 polarization, based on single-cell RNA sequencing, RNA-seq and ATAC-seq analysis.

In the repair of maxillofacial bone defects, autogenous craniofacial bone can often provide superior clinical results over long bone grafts. Most current studies have focused on the osteogenic differences between alveolar bone marrow (ABM) and long bone marrow (LBM), however, studies about the angiogenic differences between the two are currently lacking. We downloaded single-cell RNA sequencing (scRNA-seq) of mouse ABM and LBM respectively from the public database, and the data were processed by using Seurat package. CellphoneDB2 results showed that macrophages had the strongest interaction with mesenchymal stem cells (MSCs) and endothelial cells (ECs). ELISA results confirmed that ABM macrophages secreted a higher level of vascular endothelial growth factor A (Vegfa) compared to LBM macrophages, which further promoted angiogenesis of ECs and MSCs. Using SCENIC package, six key transcription factors (TFs) were identified to regulate the difference between ABM and LBM macrophages, and activating transcription factor 4 (Atf4) was confirmed to be more expressed in ABM macrophages by polymerase chain reaction (PCR) and western blot (WB), with predicted target genes including Vegfa. Besides, the result of scRNA-seq implied ABM macrophages more in M1 status than LBM macrophages, which was confirmed by the following experiments. From the results of another assay for transposase accessible chromatin sequencing (ATAC-seq) and RNA-seq about M1 macrophages, Atf4 was also confirmed to regulate the M1 polarization. So, we suspected that Atf4 regulated the different expression of Vegfa between ABM and LBM macrophages by activating M1 polarization. After knocking down Atf4, the expression of M1 polarization markers and Vegfa were downregulated and vasculogenic differences were eliminated, which were subsequently reversed by the addition of LPS/IFN-γ. Our study might provide a new idea to improve the success rate of autologous bone grafting and treatment of oral diseases.

Inhibition of CDKL3 downregulates STAT1 thus suppressing prostate cancer development.

Prostate cancer poses a great threat to men's health worldwide, yet its treatment is still limited by the unclear understanding of its molecular mechanisms. CDKL3 is a molecule with a recently discovered regulatory role in human tumors, and its relationship with prostate cancer is unknown. The outcomes of this work showed that CDKL3 was significantly upregulated in prostate cancer tissues compared with adjacent normal tissues, and was significantly positively correlated with tumor malignancy. Knockdown of CDKL3 levels in prostate cancer cells significantly inhibited cell growth and migration and enhanced apoptosis and G2 arrest of the cell cycle. Cells with lower CDKL3 expression also had relatively weaker in vivo tumorigenic capacity as well as growth capacity. Exploration of downstream mechanisms of CDKL3 may regulate STAT1, which has co-expression characteristics with CDKL3, by inhibiting CBL-mediated ubiquitination of STAT1. Functionally, STAT1 is aberrantly overexpressed in prostate cancer and has a tumor-promoting effect similar to that of CDKL3. More importantly, the phenotypic changes of prostate cancer cells induced by CDKL3 were dependent on ERK pathway and STAT1. In summary, this work identifies CDKL3 as a new prostate cancer-promoting factor, which also has the potential to be a therapeutic target for prostate cancer.

Piezo1-mediated M2 macrophage mechanotransduction enhances bone formation through secretion and activation of transforming growth factor-ß1.

Macrophages are multifunctional immune system cells that are essential for the mechanical stimulation-induced control of metabolism. Piezo1 is a non-selective calcium channel expressed in multifarious tissues to convey mechanical signals. Here, a cellular model of tension was used to study the effect of mechanical stretch on the phenotypic transformation of macrophages and its mechanism. An indirect co-culture system was used to explore the effect of macrophage activation on bone marrow mesenchymal stem cells (BMSCs), and a treadmill running model was used to validate the mechanism in vivo for in vitro studies. p53 was acetylated and deacetylated by macrophages as a result of mechanical strain being detected by Piezo1. This process is able to polarize macrophages towards M2 and secretes transforming growth factor-beta (TGF-ß1), which subsequently stimulates BMSCs migration, proliferation and osteogenic differentiation. Knockdown of Piezo1 inhibits the conversion of macrophages to the reparative phenotype, thereby affecting bone remodelling. Blockade of TGF-ß I, II receptors and Piezo1 significantly reduced exercise-increased bone mass in mice. In conclusion, we showed that mechanical tension causes calcium influx, p53 deacetylation, macrophage polarization towards M2 and TGF-ß1 release through Piezo1. These events support BMSC osteogenesis.