Single-cell transcriptomics of hepatic stellate cells uncover crucial pathways and key regulators involved in non-alcoholic steatohepatitis

Background: Fibrosis is an important pathological process in the development of non-alcoholic steatohepatitis (NASH), and the activation of hepatic stellate cell (HSC) is a central event in liver fibrosis. However, the transcriptomic change of activated HSCs (aHSCs) and resting HSCs (rHSCs) in NASH patients has not been assessed. This study aimed to identify transcriptomic signature of HSCs during the development of NASH and the underlying key functional pathways. Methods: NASH-associated transcriptomic change of HSCs was defined by single-cell RNA-sequencing (scRNA-seq) analysis, and those top upregulated genes were identified as NASH-associated transcriptomic signatures. Those functional pathways involved in the NASH-associated transcriptomic change of aHSCs were explored by weighted gene co-expression network analysis (WGCNA) and functional enrichment analyses. Key regulators were explored by upstream regulator analysis and transcription factor enrichment analysis. Results: scRNA-seq analysis identified numerous differentially expressed genes in both rHSCs and aHSCs between NASH patients and healthy controls. Both scRNA-seq analysis and in-vivo experiments showed the existence of rHSCs (mainly expressing a-SMA) in the normal liver and the increased aHSCs (mainly expressing collagen 1) in the fibrosis liver tissues. NASH-associated transcriptomic signature of rHSC (NASHrHSCsignature) and NASH-associated transcriptomic signature of aHSC (NASHaHSCsignature) were identified. WGCNA revealed the main pathways correlated with the transcriptomic change of aHSCs. Several key upstream regulators and transcription factors for determining the functional change of aHSCs in NASH were identified. Conclusion: This study developed a useful transcriptomic signature with the potential in assessing fibrosis severity in the development of NASH. This study also identified the main pathways in the activation of HSCs during the development of NASH

Delving into the role of creativity on meaning in life: A multiple mediation model

How to enhance the sense of meaning in life is a topic deserving of extensive research. The impact of creativity on finding meaning in life, however, has not been thoroughly explored in empirical research. This paper studies the relationship between creativity and meaning in life, and the cognitive and emotional factors underlying this relationship. The participants of this study were 359 Chinese college students (38 males and 321 females; aged from 17 to 41 years) in learning English as a foreign language (EFL). Four instruments were utilized in the survey, namely, the Kaufman Domains of Creativity Scale (K-DOCS), the Positive Affect Scale, the General Self-Efficacy Scale (GSES), and the Meaning in Life Questionnaire (MLQ). The correlation analysis shows that creativity, positive affect, general self-efficacy, and meaning in life are all positively correlated. According to a bootstrap method to assess the significance of the indirect effect, general self-efficacy and positive affect play multiple mediating roles in the relationship between creativity and meaning in life via three mediating pathways: general self-efficacy alone, positive affect alone, and the effect of general self-efficacy on positive affect. The mediating effect accounts for nearly half (44.18%) of the total effect. This study examines the theoretical connection between creativity and meaning in life, and uncovers the psychological process that underlies this connection. On a practical level, these results indicate that stimulating Chinese college students to engage in creative activities in various fields can enhance their sense of meaning in life

Calculation model and influence factors of thermal conductivity of composite cement-based materials for geothermal well

Abstract The use of cement-based composites (CBC) with high thermal conductivity for geothermal well cementing is extremely important for the efficient development and use of geothermal energy. Accurate prediction of thermal conductivity can save a lot of experimental costs and time. At present, there is no specific calculation model for the thermal conductivity of CBC. In this study, the microstructure, thermal conductivity model and influencing factors of CBC were investigated by experimental tests, theoretical analysis and numerical simulation. The results showed that the cement-based material could be simplified into a two-layer structure of hydrated and unhydrated layers. Mathematical and numerical models based on the coupled Series model and the Maxwell–Eucken model were established to calculate the thermal conductivity for CBC. The mathematical and numerical models were found to be more accurate by comparison with the conventional models and experimental test results. The cubic packing was more favorable than the spherical packing to improve the thermal conductivity of CBC. The plate material had significant anisotropy. The thermal conductivity of CBC showed a rapid decrease followed by a slow decrease, a decrease followed by a slow increase and finally a rapid decrease, a rapid increase followed by an up and down fluctuation and finally a plateau, respectively, with the increase of filler particle diameter, spacing and curing temperature. Based on these results, the effective methods and future research directions were proposed to maximize the thermal conductivity of geothermal well cementing materials in actual engineering applications. The research findings can provide some technical references for the efficient development of geothermal energy and research on CBC with high thermal conductivity

TSPO Deficiency Exacerbates GSDMD-Mediated Macrophage Pyroptosis in Inflammatory Bowel Disease

Background: the 18-kDa translocator protein (TSPO) is a mitochondrial outer membrane protein, and its expression tends to increase in response to inflammatory stimulation, rapidly. However, the role of TSPO in inflammation and pyroptosis is not yet clear. Here, we identified TSPO as a novel key regulator of pyroptosis. (2) Methods: TSPO knockout and DSS induced mouse inflammatory bowel disease (IBD) models were employed to assess the roles of TSPO in the pathogenesis of IBD. Primary peritoneal macrophages from TSPO knockout mice were applied to evaluate the mechanism of TSPO in cell pyroptosis. Conclusions: in response to inflammatory injury, TSPO expression is rapidly upregulated and provides a protective function against GSDMD-mediated pyroptosis, which helps us better understand the biological role of TSPO and a novel regulatory mechanism of the pyroptosis process

Scandoside Exerts Anti-Inflammatory Effect Via Suppressing NF-κB and MAPK Signaling Pathways in LPS-Induced RAW 264.7 Macrophages

The iridoids of Hedyotis diffusa Willd play an important role in the anti-inflammatory process, but the specific iridoid with anti-inflammatory effect and its mechanism has not be thoroughly studied. An iridoid compound named scandoside (SCA) was isolated from H. diffusa and its anti-inflammatory effect was investigated in lipopolysaccharide (LPS)-induced RAW 264.7 macrophages. Its anti-inflammatory mechanism was confirmed by in intro experiments and molecular docking analyses. As results, SCA significantly decreased the productions of nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) and inhibited the levels of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), TNF-α and IL-6 messenger RNA (mRNA) expression in LPS-induced RAW 264.7 macrophages. SCA treatment suppressed the phosphorylation of inhibitor of nuclear transcription factor kappa-B alpaha (IκB-α), p38, extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK). The docking data suggested that SCA had great binding abilities to COX-2, iNOS and IκB. Taken together, the results indicated that the anti-inflammatory effect of SCA is due to inhibition of pro-inflammatory cytokines and mediators via suppressing the nuclear transcription factor kappa-B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways, which provided useful information for its application and development

Identifying a distinct fibrosis subset of NAFLD via molecular profiling and the involvement of profibrotic macrophages.

BACKGROUND: There are emerging studies suggesting that non-alcoholic fatty liver disease (NAFLD) is a heterogeneous disease with multiple etiologies and molecular phenotypes. Fibrosis is the key process in NAFLD progression. In this study, we aimed to explore molecular phenotypes of NAFLD with a particular focus on the fibrosis phenotype and also aimed to explore the changes of macrophage subsets in the fibrosis subset of NAFLD. METHODS: To assess the transcriptomic alterations of key factors in NAFLD and fibrosis progression, we included 14 different transcriptomic datasets of liver tissues. In addition, two single-cell RNA sequencing (scRNA-seq) datasets were included to construct transcriptomic signatures that could represent specific cells. To explore the molecular subsets of fibrosis in NAFLD based on the transcriptomic features, we used a high-quality RNA-sequencing (RNA-seq) dataset of liver tissues from patients with NAFLD. Non-negative matrix factorization (NMF) was used to analyze the molecular subsets of NAFLD based on the gene set variation analysis (GSVA) enrichment scores of key molecule features in liver tissues. RESULTS: The key transcriptomic signatures on NAFLD including non-alcoholic steatohepatitis (NASH) signature, fibrosis signature, non-alcoholic fatty liver (NAFL) signature, liver aging signature and TGF-β signature were constructed by liver transcriptome datasets. We analyzed two liver scRNA-seq datasets and constructed cell type-specific transcriptomic signatures based on the genes that were highly expressed in each cell subset. We analyzed the molecular subsets of NAFLD by NMF and categorized four main subsets of NAFLD. Cluster 4 subset is mainly characterized by liver fibrosis. Patients with Cluster 4 subset have more advanced liver fibrosis than patients with other subsets, or may have a high risk of liver fibrosis progression. Furthermore, we identified two key monocyte-macrophage subsets which were both significantly correlated with the progression of liver fibrosis in NAFLD patients. CONCLUSION: Our study revealed the molecular subtypes of NAFLD by integrating key information from transcriptomic expression profiling and liver microenvironment, and identified a novel and distinct fibrosis subset of NAFLD. The fibrosis subset is significantly correlated with the profibrotic macrophages and M2 macrophage subset. These two liver macrophage subsets may be important players in the progression of liver fibrosis of NAFLD patients

Exploring the Relationship of Bone Turnover Markers and Bone Mineral Density in Community-Dwelling Postmenopausal Women

Aims. To explore the relationships of procollagen type 1 N-terminal propeptide (P1NP) and β cross-linked C-telopeptide of type 1 collagen (β-CTX) with bone mineral density (BMD) in postmenopausal women. Methods. All postmenopausal women were selected from a community-based case-control study. The anteroposterior L1-L4 and left proximal femur BMD were measured. P1NP and β-CTX were also collected and tested. The main correlation analysis was applied to explore the relationships of BMD, P1NP, and β-CTX. Results. The total 1055 postmenopausal women were enrolled. The BMD at all sites kept a decrease continually with age (P<0.01). In addition, the level of β-CTX increased significantly from 45 to 50 years old and remained at a high level in the later stage, while the level of P1NP changed little or even decreased with age. Logistic regression model showed that β-CTX has better ability to predict BMD than P1NP, as demonstrated by an area under the curve (AUC) of 0.63. Conclusion. P1NP and β-CTX are important markers to monitor bone metabolism. This trial is registered with ChiCTR-SOC-17013090. The date of registration is Oct. 23, 2017

ATPase Thorase Deficiency Causes &alpha;-Synucleinopathy and Parkinson&rsquo;s Disease-like Behavior

Parkinson&rsquo;s disease (PD) is one of the most common neurodegenerative diseases and is pathologically characterized by &alpha;-synucleinopathy, which is harmful to dopaminergic neurons. However, the underlying mechanisms and pathogenesis of PD remain unclear. The AAA + ATPase Thorase was identified as being essential for neuroprotection and synaptic plasticity by regulating the AMPA receptor trafficking. Here, we found that conditional knockout of Thorase resulted in motor behaviors indicative of neurodegeneration. Genetic deletion of Thorase exacerbated phenotypes of &alpha;-synucleinopathy in a familial PD-like A53T mouse model, whereas overexpression of Thorase prevented &alpha;-syn accumulation in vivo. Biochemical and cell cultures studies presented here suggest that Thorase interacts with &alpha;-syn and regulates the degradation of ubiquitinated &alpha;-syn. Thorase deficiency promotes &alpha;-syn aggregation in primary cultured neurons. The discoveries in this study provide us with a further understanding of the pathogenesis of &alpha;-synucleinopathies including PD

Genetic Deletion of Thorase Causes Purkinje Cell Loss and Impaired Motor Coordination Behavior

Thorase belongs to the AAA+ ATPase family, which plays a critical role in maintaining cellular homeostasis. Our previous work reported that Thorase was highly expressed in brain tissue, especially in the cerebellum. However, the roles of Thorase in the cerebellum have still not been characterized. In this study, we generated conditional knockout mice (cKO) with Thorase deletion in Purkinje cells. Thorase cKO mice exhibited cerebellar degenerative diseases-like behavior and significant impairment in motor coordination. Thorase deletion resulted in more Purkinje neuron apoptosis, leading to Purkinje cell loss in the cerebellum of Thorase cKO mice. We also found enhanced expression of the inflammatory protein ASC, IL-1β, IL-6 and TNF-α in the Thorase cKO cerebellum, which contributed to the pathogenesis of cerebellar degenerative disease. Our findings provide a better understanding of the role of Thorase in the cerebellum, which is a theoretical basis for Thorase as a therapeutic drug target for neurodegenerative diseases