Aspirin-triggered Resolvin D1 ameliorates activation of the NLRP3 inflammasome via induction of autophagy in a rat model of neuropathic pain.

Background: Several studies performed thus far indicate that neuroinflammation may be one of the mechanisms underlying the pathogenesis of neuropathic pain (NP). Autophagy, as an adaptive response, has been regarded as an active process of removing the inflammatory stimulus and restoring homeostatic balance. Resolution of inflammation is a biochemical process mediated by the so-called aspirin-triggered specialized proresolving lipid mediators (AT-SPMs), which are thought to exert protective effects in NP. Recent studies have proposed mechanisms in models of inflammatory disorders and showed a relationship between resolution of inflammation and autophagy. This study aimed to validate the functional effects of Aspirin-triggered Resolvin D1 (AT-RvD1) on in vitro and in vivo models of inflammation and to determine their roles in the regulation of autophagy and activation of the Nod-like receptor protein 3 (NLRP3) inflammasome signaling pathway. Methods: An NP model was established using L5-6 spinal nerve ligation (SNL) and a model of tumor necrosis factor alpha (TNF-α)-stimulated primary microglia was established to evaluate the effect of SPMs. Western blotting was used to detect the level of NLRP3 inflammasomes complexes proteins (NLRP3, ASC, and Caspase-1) and autophagy-related proteins (LC3B, and Beclin1). Immunofluorescence staining was used to understand the autophagy and NLRP3 inflammasome activation process. The behavioral changes in rats were analyzed using paw withdrawal thresholds (PWT) and paw withdrawal latency (PWL) test. Results: Our results showed that AT-SPMs significantly upregulated the activation of autophagy, which was characterized by an increase in the ratio of LC3B-II/I and accumulation of ATG5 and Beclin1. AT-RvD1 showed a dose-dependent decrease in the upregulated PWT and PWL induced by SNL and suppressed the expression of the NLRP3 inflammasome protein and the production of its corresponding downstream proinflammatory factors. Additionally, AT-RvD1 induced the activation of autophagy of the microglia and decreased the expression of the NLRP3 inflammasome protein and the accumulation of proinflammatory factors in TNF-ɑ-challenged microglia. Conclusion: Thus, these results showed that AT-RvD1 may be a potential alternative therapeutic strategy for the prevention or treatment of NP by inhibition of the NLRP3 inflammasome signaling pathway by targeting the induction of autophagy.

Does Primary Tumor Resection Induce Accelerated Metastasis in Breast Cancer? A Review.

INTRODUCTION: Over a century of cumulative experimental results and clinical data have suggested that surgical procedures of primary tumors promote tumor progression and metastasis in breast cancer and other cancer patients, suggesting a potential interplay linking primary tumors and distant lesions that lead to metastasis development triggered by primary tumor removal. Such evidence may generate a departure in terms of our attitude toward the surgery. However, the reliability and prognostic benefits of tumor surgery, especially for chemotherapy-resistant patients, are indisputable. Thus, it is important to explore the mechanism underlying this surgery-induced cancer progression to guide individual clinical treatment and improve tumor control. MATERIALS AND METHODS: We conducted a comprehensive review in PubMed in October 2021 to determine the article outline. Non-English and repetitive articles were excluded. The year, topic, key findings, and opinions of each article were gathered. RESULTS: This review not only comprehensively summarizes the potential mechanisms of primary tumors interacting with the growth of metastases but also discusses whether and how surgical resection of primary lesions can trigger tumor metastasis and development. At the same time, this article also provides our understanding of clinical findings and future directions on this topic. In addition, the combination of surgery and some potentially beneficial therapeutic interventions for postoperative tumor metastasis control was also mentioned. CONCLUSIONS: There are viewpoints supporting an acceleration of metastasis after surgery for breast cancer and fundamental research on relevant therapies, although controversial. Further attention should be focused on the gap between current preclinical data and the complicated clinical therapeutic combination during surgery in metastatic breast cancer patients.

LncRNA RASSF8-AS1 knockdown displayed antiproliferative and proapoptotic effects through miR-188-3p/ATG7 pathway in ox-LDL-treated vascular smooth muscle cells.

Background: Long noncoding RNA (lncRNA)-mediated changes in gene expression contribute to atherosclerosis (AS) development. However, the roles of numerous lncRNAs in AS have not been fully elucidated. Here, we aimed to investigate the potential role of lncRNA RASSF8-AS1 (RASSF8-AS1) in autophagy of human aortic vascular smooth muscle cells (HA-VSMCs). Methods: RASSF8-AS1 expression in patients with AS was extracted from the Gene Expression Omnibus (GEO) database. RASSF8-AS1 and microRNA-188-3p (miR-188-3p) expression was analyzed in 20 enrolled patients with AS. HA-VSMCs were treated with oxidized low-density lipoprotein (ox-LDL) (25, 50, 75, and 100 µg/mL) for 24 h. Loss- or gain-of-function of RASSF8-AS1, miR-1883p, and autophagy-related 7 (ATG7) was studied using the transfected HA-VSMCs. Cell viability was assessed using Cell Counting Kit-8 (CCK-8). Apoptosis was detected with annexin V-fluorescein isothiocyanate (FITC) and propidium iodide (PI). Relative luciferase reporter assay was used to confirm the targeting relationship of miR-188-3p to RASSF8-AS1 or ATG7. Gene expression was detected by quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot. Results: RASSF8-AS1 was enriched in the serum of patients with AS and ox-LDL-treated HA-VSMCs. Ox-LDL induced proliferation and autophagy while inhibiting the apoptosis of HA-VSMCs, which was abated by RASSF8-AS1 knockdown. RASSF8-AS1 downregulated miR-188-3p of ox-LDL-treated HA-VSMCs. RASSF8-AS1 knockdown caused an increase in miR-188-3p, which inhibited proliferation and autophagy and induced the apoptosis of ox-LDL-treated HA-VSMCs. miR-188-3p inhibited ATG7 expression in ox-LDL-treated HA-VSMCs. RASSF8-AS1 elevated ATG7 and induced autophagy through sponging miR-188-3p in ox-LDL-treated HA-VSMCs. Conclusions: RASSF8-AS1 regulated autophagy by targeting miR-188-3p, a messenger RNA-binding miRNA that increases ATG7 level, which may be a new target molecule for the prevention and prognosis of AS.

Non-hepatitis-associated mixed cryoglobulinemia with polyclonal plasma cells disease and alcoholic cirrhosis: A rare case report.

Mixed cryoglobulinemia refers to the serum presence of a variety of cryoglobulins, which are defined as immunoglobulins that precipitate at temperatures of < 37°C. The most common cause of mixed cryoglobulinemia is hepatitis C virus (HCV), while other infections, including hepatitis B virus (HBV) and HIV infections, and lymphoproliferative and autoimmune disorders have also been associated with the disease. We reported a rare case of type II-III mixed cryoglobulinemia caused by alcoholic cirrhosis. We need to increase the awareness of and facilitate the early identification of mixed cryoglobulinemia in our clinical study when encountering a patient with liver cirrhosis combined with renal impairment so that treatment can begin early to improve the success rate of therapy and reduce the fatality rate in a potentially life-saving therapy.

Breast cancer cells interact with tumor-derived extracellular matrix in a molecular subtype-specific manner.

Mimicking the native microenvironment is vital for tumor engineering. Breast cancer is a highly heterogeneous disease with various molecular subtypes exhibiting distinct biological behaviors and treatment responsiveness. The heterogeneity of extracellular matrix (ECM) of breast cancer has remained largely unexplored and underestimated. The present study addressed this issue by comparing the composition, architecture, and functional roles of ECMs derived from breast cancers of two molecular subtypes, which are luminal-A breast cancer (less aggressive, ERα+)-derived ECM (LA-ECM) and triple-negative breast cancer (high aggressive, ERα-)-derived ECM (TN-ECM). Compared with normal breast tissue-derived ECMs (B-ECM), tumor-derived ECMs showed higher contents of pro-collagen I, fibronectin, and laminin, in addition with a significantly altered architecture. Transcriptome sequencing revealed that, compared with those cultured with B-ECM, MCF7 cells (an estrogen receptor (ER)α + luminal-A breast cancer cell line) cultured with LA-ECM and TN-ECM showed approximately 9.65 % and 9.04 % changes in the expression of all detected genes, respectively. The TN-ECM induced proliferation, promoted epithelial-to-mesenchymal transition, downregulated ERα expression, and reduced endocrine treatment sensitivity of MCF7. Above results have elucidated the role of phenotype-specific tumor ECM in cell phenotype maintenance, treatment sensitivity, and cancer progression, which highlighted the importance of ECM heterogeneity as well as its role in tumor microenvironment engineering and drug screening.

Application of validated migration models for the risk assessment of styrene and acrylonitrile in ABS plastic toys.

With styrene and acrylonitrile in ABS plastic toys as examples, this paper introduces to the development of a systematic strategy for studying the chemical migration risk in toys. The approach, included the detection method, establishment of migration model, model verification, and the practical application of the model in risk assessment. First, simple and sensitive methods for detecting analyte residues and migration were developed by headspace GC-MS. Then, the migration models were established based on the migration data from 5 min to 168 h and verified using 11 ABS samples. The results showed that the predicted values of the models and the experimental values had a good fit (RMSE=0.10-8.72 %). Subsequently, the migration of analytes in 94 ABS toys was predicted with these models at specific migration times. The daily average exposure level to styrene and acrylonitrile were estimated for children (3 months to 3 years). At last, the migration models reasonably predicted that the cancer risk of styrene and acrylonitrile in ABS toys were 1.6 × 10-8-1.4 × 10-6 and 3.1 × 10-8-1.6 × 10-6, respectively. This research contributes to promote toy safety and child health by enriching migration models and risk assessments.

Prognostic Role of Serum Soluble Tim-3 in Severe Traumatic Brain Injury: A Prospective Observational Study.

Objective: T cell immunoglobulin and mucin domain-3 (Tim-3) may be implicated in neuroinflammation. Herein, we attempted to discern the role of serum soluble (s) Tim-3 as an inflammatory prognostic biomarker of severe traumatic brain injury (sTBI). Methods: In this prospective observational study of 112 sTBI patients and 112 controls, serum sTim-3 levels were determined, Rotterdam computed tomography (CT) classification and Glasgow coma scale (GCS) were selected as the two severity indicators, serum C-reactive protein (CRP) was regarded as an inflammatory biomarker, and poor prognosis was referred to as extended Glasgow outcome scale (GOSE) scores 1-4 at 180 days after trauma. Results: Serum sTim-3 levels were markedly higher in patients than in controls (median, 4.2 ng/mL versus 0.7 ng/mL; P<0.001). Serum sTim-3 levels of patients were independently related to Rotterdam CT scores (ß=1.126), GCS scores (ß=-0.589), serum CRP levels (ß=0.155) and GOSE scores (ß=-0.211). Serum sTim-3 appeared as an independent predictor of post-traumatic 180-day mortality (odds ratio=1.289), overall survival (hazard ratio=1.208) and poor prognosis (odds ratio=1.293). Serum sTim-3 levels discriminated patients at risk of post-injury 180-day mortality and poor prognosis with areas under curve (AUCs) at 0.753 and 0.782, respectively. Serum sTim-3 levels combined with GCS scores and Rotterdam CT scores (AUC=0.869) exhibited significantly higher AUC than Rotterdam CT scores (P=0.026), but not than GCS scores (P=0.181) for death prediction and their combination (AUC=0.895) had significantly higher AUC than GCS scores (P=0.036) or Rotterdam CT scores (P=0.005) for outcome prediction. Conclusion: Elevated serum sTim-3 levels, in close correlation with traumatic severity and inflammation, are substantially associated with long-term death and poor outcome, indicating that serum sTim-3, as an inflammatory biomarker, may be of clinical significance in severity assessment and prediction of prognosis following sTBI.

Enhanced Local Delivery of microRNA-145a-5P into Mouse Aorta via Ultrasound-Targeted Microbubble Destruction Inhibits Atherosclerotic Plaque Formation.

Abnormal proliferation and migration of vascular smooth muscle cells (VSMCs) play a key role in the formation and rupture of atherosclerotic plaques. Previous studies have confirmed that microRNA-145 (miR-145) is involved in the phenotypic regulation of VSMCs and reduction of atherosclerosis. At present, seeking safe and effective gene delivery remains a key problem restricting the development of gene therapy. In recent years, ultrasound-targeted microbubble destruction (UTMD) has become a safe and effective transfection method that is widely used in the basic research of gene therapy for heart and tumor diseases. Here, we synthesized cationic microbubbles to encapsulate miR-145 and targeted their release into VSMCs in vitro and in vivo using ultrasound. The feasibility of this gene therapy was verified by fluorescence microscopy and an in vivo imaging system. The results showed that treatment with miR-145 delivered via UTMD considerably improved the gene transfection efficiency and promoted the contraction phenotype of VSMCs in vitro. In vivo, this treatment reduced the atherosclerotic plaque area by 48.04% compared with treatment with free miR-145. Therefore, UTMD-mediated miRNA therapy may provide a new targeted therapeutic approach for atherosclerotic plaques.

Description of Gut Mycobiota Composition and Diversity of Caprinae Animals.

The fungal community, also known as mycobiota, plays pivotal roles in host nutrition and metabolism and has potential to cause disease. However, knowledge of the gut fungal structure in Caprinae is quite limited. In this study, the composition and diversity of the gut mycobiota of Caprinae animals from different geographical locations (Anhui, Jilin, Guangxi, Shandong, Shanxi, and Tibet) were comprehensively characterized by analyzing the internal transcribed spacer 2 (ITS-2) sequences of the fungal community. The results showed that Ascomycota and Basidiomycota were the dominant phyla, which, respectively, accounted for 90.86 to 95.27% and 2.58 to 7.62% of sequences in samples from each region. Nonetheless, the structure of the gut mycobiota was largely different in Caprinae animals in the different provinces. Therein, Sporormiaceae and Thelebolaceae were the dominant fungal families in the samples from Tibet, whereas their abundance was generally low in other regions. The intestinal diversity of individuals from Guangxi was higher than that in other regions. In addition, there were 114 differential genera among all regions. Finally, the co-occurrence network revealed 285 significant correlations in cross-family pairs in the guts of Caprinae animals, which contained 149 positive and 136 negative relationships, with 96 bacterial and 86 fungal participants at the family level. This study has improved the understanding of the mycobiota of ruminants and provided support for the improvement in animal health and productivity. IMPORTANCE In this study, we elucidated and analyzed the structure of the gut mycobiota of Caprinae animals from different regions. This study revealed differences in the structure of the gut mycobiota among Caprinae animals from different geographical environments. Based on previous findings, correlations between fungal and bacterial communities were analyzed. This study adds to previous research that has expanded the present understanding of the gut microbiome of Caprinae animals.

RecurIndex assay as an aid for adjuvant chemotherapy decisions in HR-positive HER2-negative breast cancer patients.

Background: Adjuvant chemotherapy is a major adjuvant treatment modality for hormonal receptor (HR)-positive and HER2-negative early breast cancer, but only 2%-20% of patients derive practical benefits. How to balance its potential benefits and risks becomes a challenging clinical problem. The purpose of this study was to assess whether RecurIndex assay could serve as an aid for adjuvant chemotherapy decisions in Chinese patients with HR-positive HER2-negative early breast cancer. Methods: The tissue samples of pT1-2N0 HR-positive HER2-negative breast cancer from multiple centers were detected using RecurIndex assay, based on which the patients were assigned into low- and high-risk groups. The survival outcomes of low- and high-risk patients including those with and without adjuvant chemotherapy were compared, and the risk factors for recurrence and metastasis were identified. Results: Totally 445 patients were eligible for analysis. By contrast to high-risk patients, low-risk patients represented better 7-year recurrence-free survival (RFS), distant recurrence-free survival (DRFS) and local recurrence-free survival (LRFS) rates. For low-risk patients, no significant differences were shown between those with and without adjuvant chemotherapy in 7-year RFS, DRFS and LRFS rates. These differences were also inapparent between high-risk patients with and without adjuvant chemotherapy. The multivariate model revealed high-risk patients had a significantly elevated risk of recurrence and metastasis than those at low risk. Conclusion: HR-positive HER2-negative early breast cancer patients at low risk stratified by RecurIndex assay might be exempt from adjuvant chemotherapy. Whether adjuvant chemotherapy may derive survival benefits for high-risk patients still needs larger cohorts to verify.

High expression of B4GALT1 is associated with poor prognosis in acute myeloid leukemia.

Acute myeloid leukemia is the most prevalent type of leukemia in adults and is prone to relapse and chemoresistance, with a low long-term survival rate. Therefore, the identification of quality biomarkers constitutes an urgent unmet need. High expression of beta-1,4-galactosyltransferase 1 (B4GALT1) has been observed in several cancer types; however, its function in acute myeloid leukemia has rarely been studied. Therefore, our study obtained gene expression data from The Cancer Genome Atlas (TCGA) database to analyze the relationship between B4GALT1 and LAML. We compared the expression of B4GALT1 in LAML and healthy samples using the Wilcoxon rank-sum test. Furthermore, the association between B4GALT1 and survival rates was investigated using Kaplan-Meier analysis and Cox regression. The nomogram obtained by Cox analysis predicts the effect of B4GALT1 on the prognosis. To assess B4GALT1-related genes' enrichment pathway and function and the correlation between B4GALT1 and immune features, GO/KEGG, protein-protein interaction network, and single sample gene set enrichment analysis were used. In addition, B4GALT1-specific siRNAs were used to verify the effect of B4GALT1 on apoptosis. The results showed that B4GALT1 is overexpressed in LAML and has some reference value in the diagnostic and prognostic assessment of LAML. Moreover, functional enrichment showed that B4GALT1 and its 63 associated genes were closely associated with the negative regulation of the apoptotic signaling pathway. Silencing B4GALT1 significantly promoted apoptosis. In addition, B4GALT1 expression was positively correlated with the infiltration levels of macrophages, regulatory T-cell (Tregs), and Th17 cells; in contrast, B4GALT1 expression was negatively correlated with the infiltration levels of T helper cells, Mast cells, and NK cells. In conclusion, our study shows that B4GALT1 may play a vital role in the occurrence of LAML.

Serial changes of right ventricular function assessed by three-dimensional speckle-tracking echocardiography in clinically well adult heart transplantation patients.

PURPOSE: The present study aimed to evaluate serial changes of right ventricular (RV) function in clinically well adult heart transplantation (HT) patients using three-dimensional speckle-tracking echocardiography (3D-STE). METHODS: We included 58 adult HT patients, who were free from severe valvular insufficiency, severe coronary artery disease, acute rejection, or multiple organ transplantation, and 58 healthy controls. The healthy controls were matched by the distribution of age and sex with HT group. Conventional and three-dimensional (3D) echocardiography was performed in all HT patients at 1-, 3-, 6-, 9- and 12-months post-HT. And all the healthy controls underwent conventional and 3D echocardiography when recruited. Tricuspid annular plane systolic excursion (TAPSE), S' and RV fractional area change (RV FAC) were measured. Two-dimensional RV free wall longitudinal strain (2D-RV FWLS) was derived from two-dimensional speckle-tracking echocardiography (2D-STE). 3D RV free wall longitudinal strain (3D-RV FWLS) and RV ejection fraction (RVEF) were assessed by 3D-STE. RESULTS: TAPSE, S', RV FAC, 2D-RV FWLS, 3D-RV FWLS, and RVEF increased significantly from 1 to 6 months post-HT (P < 0.05). TAPSE, S', RV FAC and 2D-RV FWLS showed no significant changes from 6 to 12 months post-HT (P > 0.05), while 3D-RV FWLS and RVEF were still significantly increased: 3D-RV FWLS (17.9 ± 1.0% vs. 18.7 ± 1.4%, P < 0.001) and RVEF (45.9 ± 2.2% vs. 46.8 ± 2.0%, P = 0.025). By 12 months post-HT, TAPSE, S', RV FAC, 2D-RV FWLS, 3D-RV FWLS and RVEF were significantly lower than the healthy controls: TAPSE (15.1 ± 2.1 mm vs. 23.5 ± 3.0 mm, P < 0.001), s' (10.3 ± 1.9 cm/s vs. 12.9 ± 2.0 cm/s, P < 0.001), RV FAC (45.3 ± 1.8% vs. 49.2 ± 3.8%, P < 0.001), 2D-RV FWLS (19.9 ± 2.3% vs. 23.5 ± 3.8%, P < 0.001), 3D-RV FWLS (18.7 ± 1.4% vs. 22.4 ± 2.3%, P < 0.001) and RVEF (46.8 ± 2.0% vs. 49.9 ± 5.7%, P < 0.001). CONCLUSION: RV systolic function improved significantly over time in clinically well adult HT patients even up to 12 months post-HT. By 12 months post-HT, the patient's RV systolic function remained lower than the control. 3D-STE may be more suitable to assess RV systolic function in HT patients.

Prognostic value of feature-tracking right ventricular longitudinal strain in heart transplant recipients.

OBJECTIVES: The prognostic value of cardiac magnetic resonance feature tracking (CMR-FT)-derived right ventricular longitudinal strain (RVLS) post-heart transplantation has not been studied. This study aimed to evaluate the prognostic significance of CMR-FT-derived RVLS, in patients post- heart transplantation and to directly compare its value with that of conventional RV ejection fraction (RVEF). METHODS: In a cohort of consecutive heart transplantation recipients who underwent CMR for surveillance, RVLS from the free wall was measured by CMR-FT. The composite endpoint was all-cause death or major adverse cardiac events. The Cox regression model was used to examine the independent association between RVLS and the endpoint. RESULTS: A total of 96 heart transplantation recipients were retrospectively included. Over a median follow-up of 41 months, 20 recipients reached the composite endpoint. The multivariate Cox analysis showed that the model with RVLS (hazard ratio [HR]:1.334; 95% confidence interval [CI]:1.148 to 1.549; p < 0.001; Akaike information criterion [AIC] = 140, C-index = 0.831) was better in predicting adverse events than the model with RVEF (HR:0.928; 95% CI: 0.868 to 0.993; p = 0.030; AIC = 149, C-index = 0.751). Furthermore, receiver operating characteristic curves revealed that the accuracy for predicting adverse events was greater for RVLS than RVEF (area under the curve: 0.85 vs 0.76, p = 0.03). CONCLUSIONS: CMR-FT-derived RVLS is an independent predictor of adverse events in post-heart transplantation, and its predictive value was better than RVEF. Therefore, our study highlighted the importance of evaluating RVLS for risk stratification after heart transplantation. KEY POINTS: • CMR-RVLS is an independent predictor of adverse events post-heart transplantation and provides greater predictive value. • CMR-RVLS may help clinicians to risk stratification in heart transplantation recipients.

Synthetic Topological Vacua of Yang-Mills Fields in Bose-Einstein Condensates.

Topological vacua are a family of degenerate ground states of Yang-Mills fields with zero field strength but nontrivial topological structures. They play a fundamental role in particle physics and quantum field theory, but have not yet been experimentally observed. Here we report the first theoretical proposal and experimental realization of synthetic topological vacua with a cloud of atomic Bose-Einstein condensates. Our setup provides a promising platform to demonstrate the fundamental concept that a vacuum, rather than being empty, has rich spatial structures. The Hamiltonian for the vacuum of topological number n=1 is synthesized and the related Hopf index is measured. The vacuum of topological number n=2 is also realized, and we find that vacua with different topological numbers have distinctive spin textures and Hopf links. Our Letter opens up opportunities for exploring topological vacua and related long-sought-after instantons in tabletop experiments.

MYBL2 regulates de novo purine synthesis by transcriptionally activating IMPDH1 in hepatocellular carcinoma cells.

BACKGROUND: Metabolic reprogramming is a hallmark of cancer, alteration of nucleotide metabolism of hepatocellular carcinoma (HCC) is not well-understood. MYBL2 regulates cell cycle progression and hepatocarcinogenesis, its role in metabolic regulation remains elusive. PATIENTS AND METHODS: Copy number, mRNA and protein level of MYBL2 and IMPDH1 were analyzed in HCC, and correlated with patient survival. Chromatin Immunoprecipitation sequencing (Chip-seq) and Chromatin Immunoprecipitation quantitative polymerase chain reaction (ChIP-qPCR) were used to explore the relationship between MYBL2 and IMPDH1. Metabolomics were used to analyze how MYBL2 affected purine metabolism. The regulating effect of MYBL2 in HCC was further validated in vivo using xenograft models. RESULTS: The Results showed that copy-number alterations of MYBL2 occur in about 10% of human HCC. Expression of MYBL2, IMPDH1, or combination of both were significantly upregulated and associated with poor prognosis in HCC. Correlation, ChIP-seq and ChIP-qPCR analysis revealed that MYBL2 activates transcription of IMPDH1, while knock-out of MYBL2 retarded IMPDH1 expression and inhibited proliferation of HCC cells. Metabolomic analysis post knocking-out of MYBL2 demonstrated that it was essential in de novo purine synthesis, especially guanine nucleotides. In vivo analysis using xenograft tumors also revealed MYBL2 regulated purine synthesis by regulating IMPDH1, and thus, influencing tumor progression. CONCLUSION: MYBL2 is a key regulator of purine synthesis and promotes HCC progression by transcriptionally activating IMPDH1, it could be a potential candidate for targeted therapy for HCC.

AKR1C3-dependent lipid droplet formation confers hepatocellular carcinoma cell adaptability to targeted therapy.

Rationale: Increased lipid droplet (LD) formation has been linked to tumor metastasis, stemness, and chemoresistance in various types of cancer. Here, we revealed that LD formation is critical for the adaptation to sorafenib in hepatocellular carcinoma (HCC) cells. We aim to investigate the LD function and its regulatory mechanisms in HCC. Methods: The key proteins responsible for LD formation were screened by both metabolomics and proteomics in sorafenib-resistant HCC cells and further validated by immunoblotting and immunofluorescence staining. Biological function of AKR1C3 was evaluated by CRISPR/Cas9-based gene editing. Isotopic tracing analysis with deuterium3-labeled palmitate or carbon13-labeled glucose was conducted to investigate fatty acid (FA) and glucose carbon flux. Seahorse analysis was performed to assess the glycolytic flux and mitochondrial function. Selective AKR1C3 inhibitors were used to evaluate the effect of AKR1C3 inhibition on HCC tumor growth and induction of autophagy. Results: We found that long-term sorafenib treatment impairs fatty acid oxidation (FAO), leading to LD accumulation in HCC cells. Using multi-omics analysis in cultured HCC cells, we identified that aldo-keto reductase AKR1C3 is responsible for LD accumulation in HCC. Genetic loss of AKR1C3 fully depletes LD contents, navigating FA flux to phospholipids, sphingolipids, and mitochondria. Furthermore, we found that AKR1C3-dependent LD accumulation is required for mitigating sorafenib-induced mitochondrial lipotoxicity and dysfunction. Pharmacologic inhibition of AKR1C3 activity instantly induces autophagy-dependent LD catabolism, resulting in mitochondrial fission and apoptosis in sorafenib-resistant HCC clones. Notably, manipulation of AKR1C3 expression is sufficient to drive the metabolic switch between FAO and glycolysis. Conclusions: Our findings revealed that AKR1C3-dependent LD formation is critical for the adaptation to sorafenib in HCC through regulating lipid and energy homeostasis. AKR1C3-dependent LD accumulation protects HCC cells from sorafenib-induced mitochondrial lipotoxicity by regulating lipophagy. Targeting AKR1C3 might be a promising therapeutic strategy for HCC tumors.

DNMT3B and TET1 mediated DNA methylation of LATS1 regulates BC progression via hippo signaling pathway.

BACKGROUND: Large tumor suppressor gene 1 (LATS1) is one of the key proteins in the Hippo signaling pathway, which plays a role in inhibiting tumor progression, but its anti-tumor role in tumors is inhibited. Nevertheless, the specific molecular mechanism of its role in breast cancer (BC) are still unclear. METHODS: The expression of LATS1 in BC was detected by RT-qCR and WB. The biological effect of LATS1 on BC was verified in vitro and in vivo. DNA methylation sequencing and MSP were used to detect the effect of DNMT3B and TET1 on LATS1 methylation. Analysis of the effect of DNMT3B and TET1 on the biological behavior of BC cells via LATS1 by cell function experiment. RESULTS: Results indicated that the expression of LATS1 is lower in BC tumor tissues and BC cell lines. low LATS1 expression BC patients have a shorter overall survival. Increasing LATS1 expression can inhibit the proliferation, migration and invasion of BC. LATS1 found to have high DNA methylation changes in the promoter region. Moreover, DNMT3B was increased in BC tissues, and DNA demethylase TET1 was decreased in BC tissues. Furthermore, DNMT3B and TET1 regulate the methylation of LATS1, and methylation of LATS1 inactivates Hippo signaling pathway. CONCLUSION: Our results indicate that the methylation of LATS1 is regulated by DNMT3B and TET1 and regulates its protein expression and the function of Hippo signaling pathway.

A dual-readout paper-based analytical device for the simultaneous determination of hexavalent Cr and total Cr.

A paper-based analytical device (PAD) is presented with colorimetric/electrochemical dual readouts for the simultaneous sensing of total chromium (Cr) and hexavalent chromium (Cr(VI)). This device consists of a homemade three-electrode system and a patterned paper chip, integrating multiple functions including electrochemical detection, fluid driving, online oxidation, and colorimetric detection. The fiberglass filter paper with a hydrophilic microchannel was used to achieve self-driving fluidics without external equipment. One end of the microchannel was integrated with a homemade three-electrode system to achieve sample loading and electrochemical detection. The middle region on the microchannel was modified with oxidizing reagents to perform online pretreatment, and the yield of Cr(III) oxidation can reach 97.9%, ensuring reliable colorimetric detection of total Cr at another end of the microchannel modified with chromogenic agents. With this device, the signals of Cr(VI) (the signal peak at 0.29 V vs. Ag/AgCl) and total Cr can be obtained in one single injection. After optimization, the limit of detection (LOD) of Cr(VI) and total Cr were 0.01 mg L^-1 and 0.06 mg L^-1 and the linear ranges were 0.05-3.0 mg L^-1 and 0.2-3.0 mg L^-1, respectively. The relative standard deviations (RSD) of the electrochemical testing of Cr(VI) results were in a range 1.3%-8.7% (n = 3), and the RSD values of the colorimetric testing of total Cr were between 0.7-9.2% (n = 3). The device's reliability was demonstrated by performing the practical speciation of Cr in tap water, river water, and electroplating wastewater while the recoveries obtained using the present method were in the range 93.5-106%. Overall, the proposed device provides high application prospect in the on-site rapid Cr speciation.

A Catalog of over 5,000 Metagenome-Assembled Microbial Genomes from the Caprinae Gut Microbiota.

Most microbiome studies regarding the ruminant digestive tract have focused on the rumen microbiota, whereas only a few studies were performed on investigating the gut microbiota of ruminants, which limits our understanding of this important component. Herein, the gut microbiota of 30 Caprinae animals (sheep and goats) from six provinces in China was characterized using ultradeep (>100 Gbp per sample) metagenome shotgun sequencing. An inventory of Caprinae gut microbial species containing 5,046 metagenomic assembly genomes (MAGs) was constructed. Particularly, 2,530 of the genomes belonged to uncultured candidate species. These genomes largely expanded the genomic repository of the current microbes in the Caprinae gut. Several enzymes and biosynthetic gene clusters encoded by these Caprinae gut species were identified. In summary, our study extends the gut microbiota characteristics of Caprinae and provides a basis for future studies on animal production and animal health. IMPORTANCE We constructed a microbiota catalog containing 5,046 MAGs from Caprinae gut from six regions of China. Most of the MAGs do not overlap known databases and appear to be potentially new species. We also characterized the functional spectrum of these MAGs and analyzed the differences between different regions. Our study enriches the understanding of taxonomic, functional, and metabolic diversity of Caprinae gut microbiota. We are confident that the manuscript will be of utmost interest to a wide range of readers and be widely applied in future research.