InferLoop: leveraging single-cell chromatin accessibility for the signal of chromatin loop.

Deciphering cell-type-specific 3D structures of chromatin is challenging. Here, we present InferLoop, a novel method for inferring the strength of chromatin interaction using single-cell chromatin accessibility data. The workflow of InferLoop is, first, to conduct signal enhancement by grouping nearby cells into bins, and then, for each bin, leverage accessibility signals for loop signals using a newly constructed metric that is similar to the perturbation of the Pearson correlation coefficient. In this study, we have described three application scenarios of InferLoop, including the inference of cell-type-specific loop signals, the prediction of gene expression levels and the interpretation of intergenic loci. The effectiveness and superiority of InferLoop over other methods in those three scenarios are rigorously validated by using the single-cell 3D genome structure data of human brain cortex and human blood, the single-cell multi-omics data of human blood and mouse brain cortex, and the intergenic loci in the GWAS Catalog database as well as the GTEx database, respectively. In addition, InferLoop can be applied to predict loop signals of individual spots using the spatial chromatin accessibility data of mouse embryo. InferLoop is available at https://github.com/jumphone/inferloop.

ARID1A loss promotes RNA editing of CDK13 in an ADAR1-dependent manner.

BACKGROUND: ARID1A, a subunit of the SWI/SNF chromatin remodeling complex, is thought to play a significant role both in tumor suppression and tumor initiation, which is highly dependent upon context. Previous studies have suggested that ARID1A deficiency may contribute to cancer development. The specific mechanisms of whether ARID1A loss affects tumorigenesis by RNA editing remain unclear. RESULTS: Our findings indicate that the deficiency of ARID1A leads to an increase in RNA editing levels and alterations in RNA editing categories mediated by adenosine deaminases acting on RNA 1 (ADAR1). ADAR1 edits the CDK13 gene at two previously unidentified sites, namely Q113R and K117R. Given the crucial role of CDK13 as a cyclin-dependent kinase, we further observed that ADAR1 deficiency results in changes in the cell cycle. Importantly, the sensitivity of ARID1A-deficient tumor cells to SR-4835, a CDK12/CDK13 inhibitor, suggests a promising therapeutic approach for individuals with ARID1A-mutant tumors. Knockdown of ADAR1 restored the sensitivity of ARID1A deficient cells to SR-4835 treatment. CONCLUSIONS: ARID1A deficiency promotes RNA editing of CDK13 by regulating ADAR1.

FitDevo: accurate inference of single-cell developmental potential using sample-specific gene weight.

The quantification of developmental potential is critical for determining developmental stages and identifying essential molecular signatures in single-cell studies. Here, we present FitDevo, a novel method for inferring developmental potential using scRNA-seq data. The main idea of FitDevo is first to generate sample-specific gene weight (SSGW) and then infer developmental potential by calculating the correlation between SSGW and gene expression. SSGW is generated using a generalized linear model that combines sample-specific information and gene weight learned from a training dataset covering scRNA-seq data of 17 previously published datasets. We have rigorously validated FitDevo's effectiveness using a testing dataset with scRNA-seq data from 28 existing datasets and have also demonstrated its superiority over current methods. Furthermore, FitDevo's broad application scope has been illustrated using three practical scenarios: deconvolution analysis of epidermis, spatial transcriptomic data analysis of hearts and intestines, and developmental potential analysis of breast cancer. The source code and related data are available at https://github.com/jumphone/fitdevo.

Astrocytes Excessively Engulf Synapses in a Mouse Model of Alzheimer's Disease.

Synapse loss is one of the most critical features in Alzheimer's disease (AD) and correlates with cognitive decline. Astrocytes mediate synapse elimination through multiple EGF-like domains 10 (MEGF10) pathways in the developing and adult brain to build the precise neural connectivity. However, whether and how astrocytes mediate synapse loss in AD remains unknown. We here find that the phagocytic receptor MEGF10 of astrocytes is significantly increased in vivo and in vitro, which results in excessive engulfment of synapses by astrocytes in APP/PS1 mice. We also observe that the astrocytic lysosomal-associated membrane protein 1 (LAMP1) is significantly elevated, colocalized with the engulfed synaptic puncta in APP/PS1 mice, and astrocytic lysosomes contain more engulfed synaptic puncta in APP/PS1 mice relative to wild type mice. Together, our data provide evidence that astrocytes excessively engulf synapses in APP/PS1 mice, which is mediated by increased MEGF10 and activated lysosomes. The approach targeting synapse engulfment pathway in astrocytes would be a potent therapy for AD.

Electrolyte Regulation of Bio-Inspired Zincophilic Additive toward High-Performance Dendrite-Free Aqueous Zinc-Ion Batteries.

Aqueous zinc-ion batteries hold attractive potential for large-scale energy storage devices owing to their prominent electrochemical performance and high security. Nevertheless, the applications of aqueous electrolytes have generated various challenges, including uncontrolled dendrite growth and parasitic reactions, thereby deteriorating the Zn anode's stability. Herein, inspired by the superior affinity between Zn2+ and amino acid chains in the zinc finger protein, a cost-effective and green glycine additive is incorporated into aqueous electrolytes to stabilize the Zn anode. As confirmed by experimental characterizations and theoretical calculations, the glycine additives can not only reorganize the solvation sheaths of hydrated Zn2+ via partial substitution of coordinated H2 O but also preferentially adsorb onto the Zn anode, thereby significantly restraining dendrite growth and interfacial side reactions. Accordingly, the Zn anode could realize a long lifespan of over 2000 h and enhanced reversibility (98.8%) in the glycine-containing electrolyte. Furthermore, the assembled Zn||α-MnO2 full cells with glycine-modified electrolyte also delivers substantial capacity retention (82.3% after 1000 cycles at 2 A g-1 ), showing promising application prospects. This innovative bio-inspired design concept would inject new vitality into the development of aqueous electrolytes.

An All-in-One "4A Hydrogel": through First-Aid Hemostatic, Antibacterial, Antioxidant, and Angiogenic to Promoting Infected Wound Healing.

Currently used wound dressings are ineffective. Hence, there is a need to develop introduce a high-performance medicament with multiple functions including rapid hemostasis and excellent antibacterial activity to meet the growing worldwide demand for wound healing products. Here, inspired by the strong adhesion of mussels and the enzyme-mimicking activity of nanometallic biomaterials, the authors developed an injectable hydrogel to overcome multiple limitations of current wound dressings. The hydrogel is synthesized via esterification reaction between poly(vinyl alcohol) (PVA) and 3,4-dihydroxyphenylalanine (DOPA), followed by catechol-metal coordination between Cu2+ and the catechol groups of DOPA to form a PVA-DOPA-Cu (PDPC) hydrogel. The PDPC hydrogel possesses excellent tissue adhesive, antioxidative, photothermal, antibacterial, and hemostatic properties. The hydrogel rapidly and efficiently stopped bleeding under different traumatic conditions, including otherwise-lethal liver injury, high-pressure carotid artery rupture, and even fatal cardiac penetration injuries in animal models. Furthermore, it is demonstrated that the PDPC hydrogel affected high-performance wound repair and tissue regeneration by accelerating re-epithelialization, promoting collagen deposition, regulating inflammation, and contributing to vascularization. The results show that PDPC hydrogel is a promising candidate for rapid hemorrhage control and efficient wound healing in multiple clinical applications.

Mechanical loading and autophagy: A study on the BoNT-A injection-induced condylar cartilage degeneration.

Botulinum toxin A (BoNT-A) has emerged as a treatment option for temporomandibular disorder (TMD). By injecting BoNT-A into the masseter muscle, it is possible to reduce mechanical loading on the temporomandibular joint (TMJ). However, numerous prior studies have indicated excessive reduction in mechanical loading can have detrimental effects on TMJ cartilage. This study proposes that autophagy, a process influenced by mechanical loading, could play a role in BoNT-A-induced mandibular condyle cartilage degeneration. To explore this hypothesis, we employed both BoNT-A injection and an excessive biting model to induce variations in mechanical loading on the condyle cartilage of C57BL/6 mice, thereby simulating an increase and decrease in mechanical loading, respectively. Results showed a significant reduction in cartilage thickness and downregulation of Runt-related transcription factor 2 (Runx2) expression in chondrocytes following BoNT-A injection. In vitro experiments demonstrated that the reduction of Runx2 expression in chondrocytes is associated with autophagy, possibly dependent on decreased YAP expression induced by low mechanical loading. This study reveals the potential involvement of the YAP/LC3/Runx2 signaling pathway in BoNT-A mediated mandibular condylar cartilage degeneration.

pH-Responsive Fluorescent Metal-Organic Framework as a Functional Nanofiller for Smart Coatings.

A zirconium-based UiO-type UiO-66-(OH)2 metal-organic framework@carbon dot composite (Zr-MOF@CD) is synthesized through a facile solvent-free thermal method. The Zr-MOF@CD exhibits pH-responsive fluorescence behavior, which emits blue fluorescence for pH < 9 at an emission wavelength of 470 nm. At pH > 9, the fluorescence color turns from blue to yellow, with the emission behavior at 535 nm. Zr-MOF@CDs can serve as functional nanofillers in the epoxy coating for the fabrication of a smart coating, which can realize coating damage warning and metal corrosion reporting. The blue fluorescence can be observed in the area of coating damage with just a minor scratch. Once the scratch is severe enough to expose the metal substrate, the cathodic reaction of oxygen reduction in the corrosion galvanic cell causes an increased pH, where the emission of yellow fluorescence can be identified. The stable fluorescence response is free from the influence of concentration, time, temperature, and the interfering substance. Zr-MOF@CDs can also serve as nanocontainers for loading with the corrosion inhibitor and realizing the repairing of metal corrosion. The development of the smart coating with dual functions of autonomous reporting and repairing holds great potential to improve the lifetime of metals in various industrial applications.

The effects of lingual buttons, precision cuts, and patient-specific attachments during maxillary molar distalization with clear aligners: Comparison of finite element analysis.

INTRODUCTION: This study aimed to analyze the biomechanical effects of the combined use of clear aligners (CA) and auxiliaries (precision cuts, lingual buttons, and patient-specific attachments) on mesial tipping and extrusion of the premolars during maxillary molars distalization. METHODS: Three-dimensional finite element method was employed to simulate clinical scenarios of CA with different auxiliaries for molar distalization. As such, 200 g of distal force was applied to the microimplants from the notches, lingual buttons, and hooks. Orthodontic tooth movement and the hydrostatic pressure in the periodontal ligament were compared. RESULTS: Adding auxiliaries can provide the maxillary arch anchorage and promote the distal tipping of premolars and retroclination of maxillary incisors. In contrast, this effect was more pronounced in patient-specific attachment applications than in other types of auxiliaries. The independent application of the CA caused mesial tipping and extrusion of the premolar and also caused the incisor proclination. CONCLUSIONS: The anchorage loss caused by the CA alone could be alleviated with the assistance of auxiliaries. Notably, patient-specific attachments further reinforce the anchorage of the anterior arch by incorporating anchor teeth as 1 anchorage unit.

[Prediction and analysis of Q-markers of Elephantopus scaber based on its UPLC fingerprint, content determination of components, and in vitro a nti-tumor activity].

This study aimed to provide scientific evidence for predicting quality markers(Q-markers) of Elephantopus scaber by establishing UPLC fingerprint of E. scaber from different geographical origins and determining the content of 13 major components, as well as conducting in vitro anti-cancer activity investigation of the main components. The chromatographic column used was Waters CORTECS UPLC C_(18)(2.1 mm×150 mm, 1.6 µm), and the mobile phase consisted of acetonitrile and 0.1% formic acid solution(gradient elution). The column temperature was set at 30 ℃, and the flow rate was 0.2 mL·min~(-1). The injection volume was 1 µL, and the detection wavelength was 240 nm. The UPLC fingerprint of E. scaber was fitted using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 edition) to determine common peaks, evaluate similarity, identify and determine the content of major components. The CCK-8 assay was used to explore the inhibitory effect of the main components on the proliferation of lung cancer cells. The results showed that in the established UPLC fingerprint of E. scaber, 35 common peaks were identified. Thirteen major components, including neochlorogenic acid(peak 1), chlorogenic acid(peak 2), cryptochlorogenic acid(peak 3), caffeic acid(peak 4), schaftoside(peak 6), galuteolin(peak 9), isochlorogenic acid B(peak 10), isochlorogenic acid A(peak 12), isochlorogenic acid C(peak 18), deoxyelephantopin(peak 28), isodeoxyelephantopin(peak 29), isoscabertopin(peak 31), and scabertopin(peak 32) were identified and quantified, and a quantitative analysis method was established. The results of the in vitro anti-cancer activity study showed that deoxyelephantopin, isodeoxyelephantopin, isoscabertopin, and scabertopin in E. scaber exhibited inhibition rates of lung cancer cell proliferation exceeding 80% at a concentration of 10 µmol·L~(-1), higher than the positive drug paclitaxel. These results indicate that the fingerprint of E. scaber is highly characteristic, and the quantitative analysis method is accurate and stable, providing references for the research on quality standards of E. scaber. Four sesquiterpene lactones in E. scaber show significant anti-cancer activity and can serve as Q-markers for E. scaber.

Single-Cell Transcriptomic Analysis Reveals a Hepatic Stellate Cell-Activation Roadmap and Myofibroblast Origin During Liver Fibrosis in Mice.

BACKGROUND AND AIMS: HSCs and portal fibroblasts (PFs) are the major sources of collagen-producing myofibroblasts during liver fibrosis, depending on different etiologies. However, the mechanisms by which their dynamic gene expression directs the transition from the quiescent to the activated state-as well as their contributions to fibrotic myofibroblasts-remain unclear. Here, we analyze the activation of HSCs and PFs in CCL4 -induced and bile duct ligation-induced fibrosis mouse models, using single-cell RNA sequencing and lineage tracing. APPROACH AND RESULTS: We demonstrate that HSCs, rather than PFs, undergo dramatic transcriptomic changes, with the sequential activation of inflammatory, migrative, and extracellular matrix-producing programs. The data also reveal that HSCs are the exclusive source of myofibroblasts in CCL4 -treated liver, while PFs are the major source of myofibroblasts in early cholestatic liver fibrosis. Single-cell and lineage-tracing analysis also uncovers differential gene-expression features between HSCs and PFs; for example, nitric oxide receptor soluble guanylate cyclase is exclusively expressed in HSCs, but not in PFs. The soluble guanylate cyclase stimulator Riociguat potently reduced liver fibrosis in CCL4 -treated livers but showed no therapeutic efficacy in bile duct ligation livers. CONCLUSIONS: This study provides a transcriptional roadmap for the activation of HSCs during liver fibrosis and yields comprehensive evidence that the differential transcriptomic features of HSCs and PFs, along with their relative contributions to liver fibrosis of different etiologies, should be considered in developing effective antifibrotic therapeutic strategies.

Improving Reporter Gene Assay Methodology for Evaluating the Ability of Compounds to Restore P53 Activity.

Tumor suppressor protein P53 induces cycle arrest and apoptosis by mediating the transcriptional expression of its target genes. Mutations causing conformational abnormalities and post-translational modifications that promote degradation are the main reasons for the loss of P53 function in tumor cells. Reporter gene assays that can scientifically reflect the biological function can help discover the mechanism and therapeutic strategies that restore P53 function. In the reporter gene system of this work, tetracycline-inducible expression of wild-type P53 was used to provide a fully activated state as a 100% activity reference for the objective measurement of biological function. It was confirmed by RT-qPCR, cell viability assay, immunofluorescence, and Western blot analysis that the above-mentioned reporter gene system could correctly reflect the differences in biological activity between the wild-type and mutants. After that, the system was tentatively used for related mechanism research and compound activity evaluation. Through the tetracycline-induced co-expression of wild-type P53 and mutant P53 in exact proportion, it was observed that the response modes of typical transcriptional response elements (TREs) to dominant negative P53 mutation effect were not exactly the same. Compared to the relative multiple-to-solvent control, the activity percentage relative to the 100% activity reference of wild-type P53 can better reflect the actual influence of the so-called P53 mutant reactivator. Similarly, relative to the 100% activity reference, it can objectively reflect the biological effects caused by the inhibitor of P53 negative factors, such as MDM2. In conclusion, this study provides a 100% activity reference and a reliable calculation model for relevant basic research and drug development.

[Exercise regulates lipid metabolism via lipophagy and its molecular mechanisms].

Lipophagy is a kind of selective autophagy, which can selectively identify and degrade lipid droplets and plays an important role in regulating cellular lipid metabolism and maintaining intracellular lipid homeostasis. Exercise can induce lipophagy and it is also an effective means of reducing body fat. In this review, we summarized the relationship between exercise and lipophagy in the liver, pancreas, adipose tissue, and the possible molecular mechanisms to provide a new clue for the prevention and treatment of fatty liver, obesity and other related metabolic diseases by exercise.

Rutin prevents tau pathology and neuroinflammation in a mouse model of Alzheimer's disease.

BACKGROUND: Tau pathology is a hallmark of Alzheimer's disease (AD) and other tauopathies. During disease progression, abnormally phosphorylated forms of tau aggregate and accumulate into neurofibrillary tangles, leading to synapse loss, neuroinflammation, and neurodegeneration. Thus, targeting of tau pathology is expected to be a promising strategy for AD treatment. METHODS: The effect of rutin on tau aggregation was detected by thioflavin T fluorescence and transmission electron microscope imaging. The effect of rutin on tau oligomer-induced cytotoxicity was assessed by MTT assay. The effect of rutin on tau oligomer-mediated the production of IL-1ß and TNF-α in vitro was measured by ELISA. The uptake of extracellular tau by microglia was determined by immunocytochemistry. Six-month-old male Tau-P301S mice were treated with rutin or vehicle by oral administration daily for 30 days. The cognitive performance was determined using the Morris water maze test, Y-maze test, and novel object recognition test. The levels of pathological tau, gliosis, NF-kB activation, proinflammatory cytokines such as IL-1ß and TNF-α, and synaptic proteins including synaptophysin and PSD95 in the brains of the mice were evaluated by immunolabeling, immunoblotting, or ELISA. RESULTS: We showed that rutin, a natural flavonoid glycoside, inhibited tau aggregation and tau oligomer-induced cytotoxicity, lowered the production of proinflammatory cytokines, protected neuronal morphology from toxic tau oligomers, and promoted microglial uptake of extracellular tau oligomers in vitro. When applied to Tau-P301S mouse model of tauopathy, rutin reduced pathological tau levels, regulated tau hyperphosphorylation by increasing PP2A level, suppressed gliosis and neuroinflammation by downregulating NF-kB pathway, prevented microglial synapse engulfment, and rescued synapse loss in mouse brains, resulting in a significant improvement of cognition. CONCLUSION: In combination with the previously reported therapeutic effects of rutin on Aß pathology, rutin is a promising drug candidate for AD treatment based its combinatorial targeting of tau and Aß.

Mechanistic Insights into the Intercalation and Interfacial Chemistry of Mesocarbon Microbeads Anode for Potassium Ion Batteries.

Mesocarbon microbeads (MCMB) are highly desirable as anode materials for rechargeable potassium ion batteries (PIBs) due to their commercially availability, high stability and low-cost. However, their charge storage and interfacial mechanisms are still unclear. In this work, the intercalation mechanisms and the solid-electrolyte-interphase (SEI) formation of the MCMB in four different electrolytes is comprehensively studied. The MCMB anodes exhibit superior rate and cycle performances via a naked K-ions sequentially staging intercalation mechanism, realizing the complete transformation from graphite to KC8 . Whereas a solvated K-ions co-intercalation mechanism of the MCMB occurs in ether-based electrolytes, which might induce graphite exfoliation and result in unsatisfied specific capacity and capacity decay. Nevertheless, this co-intercalation behavior could be effectively suppressed by a highly concentrated electrolytes. Interfacial analyses unveil the distinct SEI components, which vary with the electrolyte chemistries. These SEI components also varies from surface to bulk and especially attention should be paid to the accurate control of the concentration of the fluoroethylene carbonate additives. This work provides a panoramic understanding of the intercalation and interfacial mechanisms on the MCMB anodes for PIBs.

Superamphiphobic Magnesium Alloys with Extraordinary Environmental Adaptability.

The application of magnesium alloys is seriously limited by their poor environmental adaptability. In this work, we report a robust superamphiphobic coating, which endows magnesium alloys with extraordinary environmental adaptability. The coating was fabricated on magnesium alloys by a facile, cost-effective, and scalable method, one-step particle-free spraying. The as-treated magnesium alloys show excellent superamphiphobicity with the static contact angles (CAs) of water, ethylene glycol, benzyl alcohol, and cyclohexanol droplets of 157.5°, 155.1°, 151.7°, and 151.3°, respectively. These samples also display small dynamic CAs (0° for water and 10° for ethylene glycol) and water super-repellency, which endow magnesium surfaces with droplet impact resistance, self-cleaning, and oil-resistance functions. The simulating environmental-adaptability tests demonstrate that the as-treated magnesium alloys can remain superamphiphobic under various mechanical, chemical, and physical damages including sand impact (⩾10 cycles), water impact (v = 4.5 m·s-1, 2 impacts·s-1, 20 h), abrasion (1.0 kPa, 50 cycles), strong acid/alkaline solution (pH = 1-14), organic solvents immersion (ethylene glycol, n-hexane, ≥48 h), high temperature (200 °C, 72 h), and ultraviolet irradiation (λ = 254 nm, 672 h). The natural environmental-adaptability tests in the acidic industrial atmosphere for 40 days further confirm the robustness of the as-treated magnesium alloys under harsh environments. This work not only provides a promising method for industrially fabricating environmental-adaptable coatings on metallic materials but also paves the way for the much wider applications of magnesium alloys.

Nanotube-decorated hierarchical tantalum scaffold promoted early osseointegration.

This study aimed to fabricate a hierarchical tantalum scaffold mimicking natural bone structure to enhance osseointegration. Porous tantalum scaffolds (p-Ta) with microgradients were fabricated by selective laser melting according to a computer-aided design model. Electrochemical anodization produced nanotubes on the p-Ta surface (p-Ta-nt). SEM verified the construction of a unique nanostructure on p-Ta-nt. Contact angle and protein adsorption measurements demonstrated that p-Ta-nt have enhanced hydrophilicity and protein absorption. MC3T3-E1 preosteoblasts showed increased filamentous pseudopods and comparable cell proliferation when cultured on p-Ta-nt. Osteogenic marker gene (Osterix, Runx2, COL-I) transcription was significantly upregulated in MC3T3-E1 cells cultured on p-Ta-nt after 7 days. After implantation into the femurs of New Zealand white rabbits for 2 weeks, histological examination found improved early osseointegration in the p-Ta-nt group. This study showed that a hierarchical tantalum structure could enhance early osteogenic effects in vitro and in vivo.

MYC-regulated lncRNA NEAT1 promotes B cell proliferation and lymphomagenesis via the miR-34b-5p-GLI1 pathway in diffuse large B-cell lymphoma.

BACKGROUND: LncRNA NEAT1 has been identified as a tumour driver in many human cancers. However, the underlying mechanism of lncRNA NEAT1 in diffuse large B-cell lymphoma (DLBCL) progression is unclear. METHODS: The expression levels of NEAT1, GLI1 and miR-34b-5p were detected by RT-qPCR and Western blotting in DLBCL tissues and cell lines. MTT and colony formation assays were performed to examine cell proliferation, while annexin-V staining and TUNEL assays were performed to measure cell apoptosis. The effect of NEAT1, GLI1 and miR-34b-5p on cell cycle-associated proteins was evaluated by Western blotting. Dual-luciferase reporter and RNA immunoprecipitation (RIP) assays were employed to investigate the interaction between NEAT1 and miR-34b-5p or GLI1 and miR-34b-5p. Moreover, chromatin immunoprecipitation (ChIP) was performed to demonstrate the interaction between MYC and NEAT1. RESULTS: NEAT1 and GLI1 were upregulated while miR-34b-5p was downregulated in DLBCL tissues and cell lines compared to normal controls. Knockdown of NEAT1 or overexpression of miR-34b-5p inhibited cell proliferation but promoted cell apoptosis. Overexpression of NEAT1 reversed GLI1-knockdown induced attenuation of cell proliferation. In other words, NEAT1 acted as a competing endogenous RNA (ceRNA), regulating the miR-34b-5p-GLI1 axis, further affecting the proliferation of DLBCL. Moreover, MYC modulated NEAT1 transcription by directly binding to the NEAT1 promoter. CONCLUSION: We revealed that MYC-regulated NEAT1 promoted DLBCL proliferation via the miR-34b-5p-GLI1 pathway, which could provide a novel therapeutic target for DLBCL.

Impact of diabetes mellitus simulations on bone cell behavior through in vitro models.

Diabetes mellitus (DM) is related to impaired bone healing and an increased risk of bone fractures. While it is recognized that osteogenic differentiation and the function of osteoblasts are suppressed in DM, the influence of DM on osteoclasts is still unclear. Hyperglycemia and inflammatory environment are the hallmark of DM that causes dysregulation of various pro-inflammatory cytokines and alternated gene expression in periodontal ligament cells, osteoblasts, osteocytes, osteoclasts, and osteoclast precursors. A methodological review on conceptual and practical implications of in vitro study models is used for DM simulation on bone cells. Several major databases were screened to find literature related to the study objective. Published literature within last 20 years that used in vitro DM-simulated models to study how DM affects the cellular behavior of bone cells were selected for this review. Studies utilizing high glucose and serum acquired from diabetic animals are the mainly used methods to simulate the diabetic condition. The combination with various simulating factors such as lipopolysaccharide (LPS), hydrogen peroxide (H2O2), and advanced glycation end products (AGEs) have been reported in diabetic situations in vitro, as well. Through screening procedure, it was evident DM-simulated conditions exerted negative impact on bone-related cells. However, inconsistent results were found among different reported studies, which could be due to variation in culture conditions, concentrations of the stimulating factors and cell lineage, etc. This manuscript has concisely reviewed currently existing DM-simulated in vitro models and provides valuable insights of detailed components in simulating DM conditions in vitro. Studies using DM-simulated microenvironment revealed that in vitro simulation negatively impacted periodontal ligament cells, osteoblasts, osteocytes, osteoclasts, and osteoclast precursors. Contrarily, studies also indicated beneficial influence on bone-related cells when such conditions are reversed.

Age-dependent association of CYP2C19 polymorphisms with clinical outcome of clopidogrel therapy in minor stroke patients with large-artery atherosclerosis.

PURPOSE: Previous studies on the association between CYP2C19 polymorphisms and therapeutic outcome of clopidogrel in stroke patients are inconclusive. We aimed to investigate the impact of CYP2C19 polymorphisms on therapeutic efficacy of clopidogrel in both young and old minor stroke patients associated with large-artery atherosclerosis (LAA). METHODS: A total of 510 eligible patients were enrolled between April 2015 and April 2018. During 1 year of follow-up, the modified Rankin Scale (mRS) was recorded. Statistical comparisons were performed using Pearson's chi squared test, Mann Whitney U test, and the Breslow-Day test to determine the effects of CYP2C19 polymorphisms on clinical outcome in different age strata. Multivariate binary logistic analysis was used to examine the potential prognostic predictors for clinical outcome. Model fitness was detected with Hosmer-Lemeshow test. RESULTS: Sixty years old was identified as the optimal cutoff age for CYP2C19 polymorphisms to affect the clinical outcome of clopidogrel therapy in LAA-associated minor stroke patients (OR = 1.67; 95% CI 1.08-2.58). Comparisons of baseline characteristics between patients with favorable and poor outcome indicated the correlation between CYP2C19 loss-of-function (LOF) allele and poorer clinical outcome in ≤ 60-year-old patients (OR = 4.29; 95% CI 1.68-10.93). The heterogeneity test showed a presence of interaction between age and CYP2C19 LOF (OR = 3.75; 95% CI 1.30-10.81). The logistic analyses further suggested that CYP2C19 LOF predicted poor clinical outcome in ≤ 60-year-old but not in > 60-year-old LAA-associated minor stroke patients receiving clopidogrel for the second prevention. CONCLUSIONS: Carriage of the CYP2C19 LOF allele may prevent expected clinical outcome during clopidogrel therapy in young LAA-associated minor stroke patients, whereas not in older patients.