Lipotoxic hepatocyte-derived exosomal miR-1297 promotes hepatic stellate cell activation through the PTEN signaling pathway in metabolic-associated fatty liver disease

BACKGROUNDExosomes play an important role in metabolic-associated fatty liver disease (MAFLD), but the mechanism by which exosomes participate in MAFLD still remain unclear.AIMTo figure out the function of lipotoxic exosomal miR-1297 in MAFLD.METHODSMicroRNA sequencing was used to detect differentially expressed miRNAs (DE-miR) in lipotoxic exosomes derived from primary hepatocytes. Bioinformatic tools were applied to analyze the target genes and pathways regulated by the DE-miRs. Quantitative real-time PCR (qPCR) was conducted for the verification of DE-miRs. qPCR, western blot, immunofluorescence staining and ethynyl-20-deoxyuridine assay were used to evaluate the function of lipotoxic exosomal miR-1297 on hepatic stellate cells (LX2 cells). A luciferase reporter experiment was performed to confirm the relationship of miR-1297 and its target gene PTEN. RESULTSMicroRNA sequencing revealed that there were 61 exosomal DE-miRs (P < 0.05) with a fold-change > 2 from palmitic acid treated primary hepatocytes compared with the vehicle control group. miR-1297 was the most highly upregulated according to the microRNA sequencing. Bioinformatic tools showed a variety of target genes and pathways regulated by these DE-miRs were related to liver fibrosis. miR-1297 was overexpressed in exosomes derived from lipotoxic hepatocytes by qPCR. Fibrosis promoting genes (α-SMA, PCNA) were altered in LX2 cells after miR-1297 overexpression or miR-1297-rich lipotoxic exosome incubation via qPCR and western blot analysis. Immunofluorescence staining and ethynyl-20-deoxyuridine staining demonstrated that the activation and proliferation of LX2 cells were also promoted after the above treatment. PTEN was found to be the target gene of miR-1297 and knocking down PTEN contributed to the activation and proliferation of LX2 cells via modulating the PI3K/AKT signaling pathway.CONCLUSIONmiR-1297 was overexpressed in exosomes derived from lipotoxic hepatocytes. The lipotoxic hepatocyte-derived exosomal miR-1297 could promote the activation and proliferation of hepatic stellate cells through the PTEN/PI3K/AKT signaling pathway, accelerating the progression of MAFLD.     

Epigenetic age estimation in saliva and in buccal cells

Age estimation based on epigenetic markers is a DNA intelligence tool with the potential to provide relevant information for criminal investigations, as well as to improve the inference of age-dependent physical characteristics such as male pattern baldness or hair color. Age prediction models have been developed based on different tissues, including saliva and buccal cells, which show different methylation patterns as they are composed of different cell populations. On many occasions in a criminal investigation, the origin of a sample or the proportion of tissues is not known with certainty, for example the provenance of cigarette butts, so use of combined models can provide lower prediction errors.In the present study, two tissue-specific and seven age-correlated CpG sites were selected from publicly available data from the Illumina HumanMethylation 450 BeadChip and bibliographic searches, to help build a tissue-dependent, and an age-prediction model, respectively. For the development of both models, a total of 184 samples (N = 91 saliva and N = 93 buccal cells) ranging from 21 to 86 years old were used. Validation of the models was performed using either k-fold cross-validation and an additional set of 184 samples (N = 93 saliva and N = 91 buccal cells, 21–86 years old).The tissue prediction model was developed using two CpG sites (HUNK and RUNX1) based on logistic regression that produced a correct classification rate for saliva and buccal swab samples of 88.59 % for the training set, and 83.69 % for the testing set. Despite these high success rates, a combined age prediction model was developed covering both saliva and buccal cells, using seven CpG sites (cg10501210, LHFPL4, ELOVL2, PDE4C, HOXC4, OTUD7A and EDARADD) based on multivariate quantile regression giving a median absolute error (MAE): ± 3.54 years and a correct classification rate ( %CP±PI) of 76.08 % for the training set, and an MAE of ± 3.66 years and a %CP±PI of 71.19 % for the testing set. The addition of tissue-of origin as a co-variate to the model was assessed, but no improvement was detected in age predictions. Finally, considering the limitations usually faced by forensic DNA analyses, the robustness of the model and the minimum recommended amount of input DNA for bisulfite conversion were evaluated, considering up to 10 ng of genomic DNA for reproducible results. The final multivariate quantile regression age predictor based on the models we developed has been placed in the open-access Snipper forensic classification website.     

miRNAs介导辐射旁效应研究进展

辐射旁效应是指受辐射的细胞产生信号,并诱导未受辐照的细胞产生反应,即受辐射和未受辐射细胞之间的通讯以及这两种细胞内的信号转导效应。辐射对肿瘤细胞造成杀伤作用的同时,也会产生辐射旁效应给邻近的正常组织带来潜在危险。研究发现,电离辐射可直接改变miRNAs表达,并影响未受辐射的邻近组织中基因的表达,因此,miRNAs可能是受辐射细胞和未受辐射细胞之间信号通路调节的重要物质。本文就miRNAs在辐射旁效应中的作用进行综述。     

Fighting viruses with materials science: Prospects for antivirus surfaces,drug delivery systems and artificial intelligence

ObjectiveViruses on environmental surfaces, in saliva and other body fluids represent risk of contamination for general population and healthcare professionals. The development of vaccines and medicines is costly and time consuming. Thus, the development of novel materials and technologies to decrease viral availability, viability, infectivity, and to improve therapeutic outcomes can positively impact the prevention and treatment of viral diseases.MethodsHerein, we discuss (a) interaction mechanisms between viruses and materials, (b) novel strategies to develop materials with antiviral properties and oral antiviral delivery systems, and (c) the potential of artificial intelligence to design and optimize preventive measures and therapeutic regimen.ResultsThe mechanisms of viral adsorption on surfaces are well characterized but no major breakthrough has become clinically available. Materials with fine-tuned physical and chemical properties have the potential to compromise viral availability and stability. Emerging strategies using oral antiviral delivery systems and artificial intelligence can decrease infectivity and improve antiviral therapies.SignificanceEmerging viral infections are concerning due to risk of mortality, as well as psychological and economic impacts. Materials science emerges for the development of novel materials and technologies to diminish viral availability, infectivity, and to enable enhanced preventive and therapeutic strategies, for the safety and well-being of humankind.     

Saliva flow rates and clinical characteristics of patients with burning mouth syndrome: A case–control study

Burning mouth syndrome (BMS) is a chronic pain condition that most commonly affects postmenopausal women older than 50 years of age. Xerostomia is a common complaint among BMS patients. However, previous studies showed inconsistent findings regarding saliva flow rate reduction. This study examined saliva flow rates, degree of mucosal hydration, xerostomia, and clinical characteristics in BMS patients compared with healthy controls. Unstimulated whole saliva (USWS) was collected through passive drooling; residual mucosal saliva (RMS) was collected using filter paper strips. Stimulated whole saliva (SWS) was collected while chewing on gum base. Oral exam and self-report data were collected. A total of 50 women (22 BMS cases and 28 healthy controls) aged 50 years or older were included in the analysis of this study. Mean age was 62 years for cases and 56 years for controls (P = 0.05). Compared with controls, cases had significantly lower USWS flow rates (P < 0.001) and had a higher prevalence of xerostomia (P = 0.001), gastrointestinal disease (P < 0.001), and vaginal dryness (P = 0.01). These data show that oral and vaginal dryness are common among BMS patients. Further studies are needed to investigate potential pathophysiological mechanisms related to the quality of saliva and mucosal barrier status among these patients.     

氟磷灰石玻璃陶瓷、长石质饰瓷与天然牙摩擦磨损性能的研究

目的：研究牙科氟磷灰石玻璃陶瓷、长石质饰瓷与天然牙在干摩擦和唾液润滑条件下的摩擦磨损学性能。方法：制备涂覆IPS-emaxcream和Vita vm9 2种饰面瓷的氧化锆-饰面瓷双层材料试样,尺寸为20 mm×20 mm×4.5 mm,各20个。利用MMV-1摩擦磨损试验机,通过设置不同大小的垂直载荷和转速,分别在干摩擦和唾液润滑条件下,对2种氧化锆-饰面瓷试样进行摩擦磨损试验。采用SPSS 17.0软件包进行单因素方差分析和t检验,确定各组间摩擦系数和磨损量是否存在差异。结果：氟磷灰石玻璃陶瓷在唾液润滑条件下的动态摩擦系数和磨损量显著大于干摩擦条件下的数值;与氟磷灰石玻璃陶瓷相反,长石质饰瓷在干摩擦时的动态摩擦系数和磨损量显著大于在唾液润滑条件下的数值。在干摩擦和唾液润滑条件下,氟磷灰石玻璃陶瓷的磨损量均小于长石质饰瓷。扫描电镜显示,在唾液润滑条件下,氟磷灰石玻璃陶瓷及对应天然牙表现出比干摩擦更为严重的磨损表面。在干摩擦条件下,长石质饰瓷及对应天然牙较氟磷灰石玻璃陶瓷磨损更为严重,在唾液润滑条件下差异较小。结论：与长石质饰瓷相比,氟磷灰石玻璃陶瓷与天然牙在干摩擦条件下表现出更优越的耐磨损性能。加入唾液后,能减少2种饰瓷摩擦磨损性能之间的差异。     

健康成人唾液与血浆中巯基化合物浓度增龄性变化初探

目的 分析健康成人唾液与血浆巯基化合物浓度与年龄、性别的关系,探讨唾液与血浆巯基化合物浓度的关联及增龄性变化,为临床检测提供参考.方法 选取2013年12月至2014年6月于解放军总医院健康管理研究院国宾部体检中心体检的健康成年志愿者306名(151名女性,155名男性),分为青年组(106名,20～ 44岁,48名女性,58名男性)、中年组(109名,45～ 59岁,63名女性,46名男性)和老年组(91名,60～ 79岁,40名女性,51名男性),分别检测唾液与血浆巯基化合物浓度.结果 306名健康成人唾液与血浆巯基化合物浓度分别为(123±27)和(427±124) μmol/L.随着年龄的增加,唾液与血浆巯基化合物浓度均显著减少(P＜0.01);老年组唾液和血浆巯基化合物浓度均显著低于青年组和中年组(P＜0.01);唾液巯基化合物浓度的男女总体差异无统计学意义(P=0.451),但血浆巯基化合物浓度的男女总体差异有统计学意义(P=0.006);唾液和血浆巯基化合物浓度显著正相关(r=0.5050,P＜O.01).结论 唾液巯基化合物浓度能大体反映血浆巯基化合物浓度,唾液巯基化合物检测有望代替血浆巯基化合物检测成为衰老的生物学指标.     

Efficacy of various cleaning solutions on saliva-contaminated zirconia for improved resin bonding

PURPOSE

This study aimed to investigate the efficacy of cleaning solutions on saliva-contaminated zirconia in comparison to air-abrasion in terms of resin bonding.

MATERIALS AND METHODS

For saliva-contaminated airabraded zirconia, seven cleaning methods)-no contamination (NC), water-spray rinsing (WS), additional airabrasion (AA), and cleaning with four solutions (Ivoclean [IC]; 1.0 wt% sodium dodecyl sulfate [SDS], 1.0 wt% hydrogen peroxide [HP], and 1.0 wt% sodium hypochlorite [SHC])-were tested. The zirconia surfaces for each group were characterized using various analytical techniques. Three bonded resin (Panavia F 2.0) cylinders (bonding area: 4.5 mm²) were made on one zirconia disk specimen using the Ultradent jig method [four disks (12 cylinders)/group; a total of 28 disks]. After 5,000 thermocycling, all specimens were subjected to a shear bond strength test with a crosshead speed of 1.0 mm/minute. The fractured surfaces were observed using an optical and scanning electron microscope (SEM).

RESULTS

Contact angle measurements showed that groups NC, AA, IC, and SHC had hydrophilic surfaces. The X-ray photoelectron spectroscopy (XPS) analysis showed similar elemental distributions between group AA and groups IC and SHC. Groups IC and SHC showed statistically similar bond strengths to groups NC and AA (P>.05), but not groups SDS and HP (P<.05). For groups WS, SDS, and HP, blister-like bubble formations were observed on the surfaces under SEM.

CONCLUSION

Within the limitations of this in vitro study, some of the cleaning solutions (IC or SHC) were effective in removing saliva contamination and enhancing the resin bond strength.