Targeting macrophagic 17ß-HSD7 by fenretinide for the treatment of nonalcoholic fatty liver disease.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease worldwide and macrophage polarization plays an important role in its pathogenesis. However, which molecule regulates macrophage polarization in NAFLD remains unclear. Herein, we showed NAFLD mice exhibited increased 17ß-hydroxysteroid dehydrogenase type 7 (17ß-HSD7) expression in hepatic macrophages concomitantly with elevated M1 polarization. Single-cell RNA sequencing on hepatic non-parenchymal cells isolated from wild-type littermates and macrophage-17ß-HSD7 knockout mice fed with high fat diet (HFD) for 6 weeks revealed that lipid metabolism pathways were notably changed. Furthermore, 17ß-HSD7 deficiency in macrophages attenuated HFD-induced hepatic steatosis, insulin resistance and liver injury. Mechanistically, 17ß-HSD7 triggered NLRP3 inflammasome activation by increasing free cholesterol content, thereby promoting M1 polarization of macrophages and the secretion of pro-inflammatory cytokines. In addition, to help demonstrate that 17ß-HSD7 is a potential drug target for NAFLD, fenretinide was screened out from an FDA-approved drug library based on its 17ß-HSD7 dehydrogenase inhibitory activity. Fenretinide dose-dependently abrogated macrophage polarization and pro-inflammatory cytokines production, and subsequently inhibited fat deposition in hepatocytes co-cultured with macrophages. In conclusion, our findings suggest that blockade of 17ß-HSD7 signaling by fenretinide would be a drug repurposing strategy for NAFLD treatment.

A dual alignment-based multi-source domain adaptation framework for motor imagery EEG classification.

Domain adaptation, as an important branch of transfer learning, can be applied to cope with data insufficiency and high subject variabilities in motor imagery electroencephalogram (MI-EEG) based brain-computer interfaces. The existing methods generally focus on aligning data and feature distribution; however, aligning each source domain with the informative samples of the target domain and seeking the most appropriate source domains to enhance the classification effect has not been considered. In this paper, we propose a dual alignment-based multi-source domain adaptation framework, denoted DAMSDAF. Based on continuous wavelet transform, all channels of MI-EEG signals are converted respectively and the generated time-frequency spectrum images are stitched to construct multi-source domains and target domain. Then, the informative samples close to the decision boundary are found in the target domain by using entropy, and they are employed to align and reassign each source domain with normalized mutual information. Furthermore, a multi-branch deep network (MBDN) is designed, and the maximum mean discrepancy is embedded in each branch to realign the specific feature distribution. Each branch is separately trained by an aligned source domain, and all the single branch transfer accuracies are arranged in descending order and utilized for weighted prediction of MBDN. Therefore, the most suitable number of source domains with top weights can be automatically determined. Extensive experiments are conducted based on 3 public MI-EEG datasets. DAMSDAF achieves the classification accuracies of 92.56%, 69.45% and 89.57%, and the statistical analysis is performed by the kappa value and t-test. Experimental results show that DAMSDAF significantly improves the transfer effects compared to the present methods, indicating that dual alignment can sufficiently use the different weighted samples and even source domains at different levels as well as realizing optimal selection of multi-source domains.

Pentoxifylline attenuates nonalcoholic fatty liver by inhibiting hepatic macrophage polarization to the M1 phenotype.

BACKGROUND: Nonalcoholic fatty liver (NAFL), recognized as one of the most common causes of chronic liver diseases, is increasingly prevalent worldwide. Pentoxifylline, a derivative of theobromine extracted from Theobroma cacao and tea, has been studied for effects on blood viscosity, tissue oxygenation and inflammation. However, its effects on hepatic lipid accumulation and the potential mechanisms remain unclear. PURPOSE: This study aimed to investigate the therapeutic effects of pentoxifylline on high-fat diet-induced NAFL and to explore the corresponding molecular mechanisms. METHODS: NAFL mice were injected with or without 25, 50 or 100 mg/kg pentoxifylline for 2 weeks. Hepatic steatosis was observed by haematoxylin-eosin staining and Oil Red O staining, the levels of serum total cholesterol, triglyceride were detected by biochemical kits, and insulin resistance was evaluated by glucose and insulin tolerance tests. In addition, we measured the frequencies of macrophage and its polarization subsets in the liver using flow cytometry and immunofluorescence. The expressions of proteins associated with macrophage polarization signaling pathways were assessed by Western blotting and flow cytometry histograms. Molecular docking and cellular thermal shift assay were conducted to identify and verify the target protein of pentoxifylline in macrophage. RESULTS: Pentoxifylline significantly alleviated hepatic lipid accumulation, reduced blood lipid levels and improved insulin resistance. Strikingly, the excessive M1 macrophages in NAFL development was abolished by pentoxifylline. And pentoxifylline was further evidenced it failed to reduce hepatocyte lipid accumulation in the absence of macrophages in vitro. Mechanistically, pentoxifylline competed with LPS for binding to toll-like receptor 4, dramatically inhibiting the TLR4/MyD88/NF-κB signaling pathway. CONCLUSION: Pentoxifylline attenuated NAFL by inhibiting hepatic macrophage M1 polarization, indicating that pentoxifylline could be a therapeutic candidate for NAFL. This study first observed that M1 macrophages were increased in NAFL mice and then revealed the molecule targeted by pentoxifylline. In addition, we provided evidence that macrophage targeting may be an emerging strategy for NAFL treatment.

[Parameter transfer learning based on shallow visual geometry group network and its application in motor imagery classification].

Transfer learning is provided with potential research value and application prospect in motor imagery electroencephalography (MI-EEG)-based brain-computer interface (BCI) rehabilitation system, and the source domain classification model and transfer strategy are the two important aspects that directly affect the performance and transfer efficiency of the target domain model. Therefore, we propose a parameter transfer learning method based on shallow visual geometry group network (PTL-sVGG). First, Pearson correlation coefficient is used to screen the subjects of the source domain, and the short-time Fourier transform is performed on the MI-EEG data of each selected subject to acquire the time-frequency spectrogram images (TFSI). Then, the architecture of VGG-16 is simplified and the block design is carried out, and the modified sVGG model is pre-trained with TFSI of source domain. Furthermore, a block-based frozen-fine-tuning transfer strategy is designed to quickly find and freeze the block with the greatest contribution to sVGG model, and the remaining blocks are fine-tuned by using TFSI of target subjects to obtain the target domain classification model. Extensive experiments are conducted based on public MI-EEG datasets, the average recognition rate and Kappa value of PTL-sVGG are 94.9% and 0.898, respectively. The results show that the subjects' optimization is beneficial to improve the model performance in source domain, and the block-based transfer strategy can enhance the transfer efficiency, realizing the rapid and effective transfer of model parameters across subjects on the datasets with different number of channels. It is beneficial to reduce the calibration time of BCI system, which promote the application of BCI technology in rehabilitation engineering.

Characterization of a broad-spectrum endolysin LysSP1 encoded by a Salmonella bacteriophage.

Foodborne pathogens have caused many public health incidents and heavy economic burden. Endolysins have been proven to have efficient bactericidal activity against pathogens with low incidence of resistance. In this study, the recombinant endolysin LysSP1 encoded by Salmonella Typhimurium lytic bacteriophage SLMP1 was obtained by prokaryotic expression, and its characteristics were analyzed. Ethylenediaminetetraacetic acid (EDTA) can be used as the outer membrane permeabilizer to increase the bactericidal activity of LysSP1. Under the synergism of 5 mmol/L EDTA, LysSP1 exhibited a strong bactericidal activity against Salmonella Typhimurium ATCC14028. LysSP1 was stable at 4°C for 7 days and at -20°C for 180 days. LysSP1 remained the optimal activity at 40°C and was efficiently active at alkaline condition (pH 8.0-10.0). Divalent metal ions could not enhance the bactericidal activity of LysSP1 and even caused the significant reduction of bactericidal activity. LysSP1 not only could lyse Salmonella, but also could lyse other Gram-negative strains and Gram-positive strains. These results indicated that LysSP1 is a broad-spectrum endolysin and has potential as an antimicrobial agent against Salmonella and other foodborne pathogens. KEY POINTS: • Recombinant endolysin LysSP1 can be prepared by prokaryotic expression. • LysSP1 has stable nature and strong bactericidal activity on Salmonella Typhimurium with EDTA. • LysSP1 has a broad range of hosts including Gram-negative bacteria and Gram-positive bacteria.

Augmented autophagy suppresses thymocytes development via Bcl10/p-p65 pathway in prenatal nicotine exposed fetal mice.

Tobacco smoke is a common global environmental pollutant. Maternal tobacco smoke/nicotine exposure has long-term toxic effects on immune organs. We previously found that prenatal nicotine exposure (PNE)-induced programmed immune diseases caused by fetal thymic hypoplasia, but the mechanism still unknown. Autophagy has important functions in maintaining thymopoiesis, whether autophagy was involved in PNE-inhibited fetal thymocytes development is also obscure. Therefore, this study aimed to investigate how nicotine changed the development of fetal thymocytes from the perspective of autophagy in vivo and in vitro. PNE model was established by 3 mg/kg nicotine administration in Balb/c mice from gestational day 9 to 18. The results showed that PNE reduced the percentage and absolute number of CD69-CD4+SP cells, suggesting a block of fetal thymocytes mature. PNE promoted autophagosome formation, autophagy related proteins (Beclin1, LC3I/II) expression, and upregulated α7 nAChR as well as AMPK phosphorylation in fetal thymus. Moreover, PNE promoted Bcl10 degradation via autophagy-mediated proteolysis and inhibited p65 activation, blocking the transition of thymocytes between the DP to SP stage. Further, primary thymocytes were treated with nicotine in vitro and showed induced autophagy in a dose- and time-dependent manner. In addition, nicotine-inhibited CD69-CD4+SP cells and the Bcl10/p-p65 pathway have been reversed by an autophagy inhibitor. The α7 nAChR specific antagonist abrogated nicotine-induced AMPK phosphorylation and autophagy initiation. In conclusion, our findings showed that PNE repressed the Bcl10/p-p65 development pathway of CD4+SP cells by triggering autophagy, and illuminated the developmental origin mechanism of programmed immune diseases in PNE offspring.

An Improved Composite Multiscale Fuzzy Entropy for Feature Extraction of MI-EEG.

Motor Imagery Electroencephalography (MI-EEG) has shown good prospects in neurorehabilitation, and the entropy-based nonlinear dynamic methods have been successfully applied to feature extraction of MI-EEG. Especially based on Multiscale Fuzzy Entropy (MFE), the fuzzy entropies of the τ coarse-grained sequences in τ scale are calculated and averaged to develop the Composite MFE (CMFE) with more feature information. However, the coarse-grained process fails to match the nonstationary characteristic of MI-EEG by a mean filtering algorithm. In this paper, CMFE is improved by assigning the different weight factors to the different sample points in the coarse-grained process, i.e., using the weighted mean filters instead of the original mean filters, which is conductive to signal filtering and feature extraction, and the resulting personalized Weighted CMFE (WCMFE) is more suitable to represent the nonstationary MI-EEG for different subjects. All the WCMFEs of multi-channel MI-EEG are fused in serial to construct the feature vector, which is evaluated by a back-propagation neural network. Based on a public dataset, extensive experiments are conducted, yielding a relatively higher classification accuracy by WCMFE, and the statistical significance is examined by two-sample t-test. The results suggest that WCMFE is superior to the other entropy-based and traditional feature extraction methods.

AMPK decreases ERK1/2 activity and cancer cell sensitivity to nutrition deprivation by mediating a positive feedback loop involving eEF2K.

Nutrition deprivation (ND) is a common feature of the tumor microenvironment. Tumor cells, therefore, frequently develop resistance mechanisms against ND. One of these mechanisms is the activation of the AMP-activated protein kinase (AMPK), which promotes cell survival under ND. AMPK activation promotes the activity of eukaryotic elongation factor 2 kinase (eEF2K), thereby blocking protein synthesis. The results of the present study indicated the inhibiting effect of AMPK activation on mitogen-activated protein kinase (ERK1/2) activity, which in turn downregulates G1/S transition and promotes cell survival by mediating eEF2K under ND. The knockdown of ERK1/2 enhances cancer cell survival under ND. In the presence of nutrients, eEF2k interacts with dual-specificity mitogen-activated protein kinase kinase (MEK)1/2, conferring a positive feedback loop via MEK1/2-ERK1/2-ribosomal protein S6 kinase α-1-eEF2K signaling, leading to the constitutive activation of ERK1/2. By contrast, under acute ND, AMPK activation blocked the interaction between eEF2K and MEK1/2, contributing to the increased resistance of cancer cells to ND. The present findings reveal a previously undiscovered mechanism that uses AMPK activation to mediate ERK1/2-regulated protein synthesis and cell survival by inhibiting eEF2K-MEK1/2 interaction under ND conditions.

Biosensors for early diagnosis of pancreatic cancer: a review.

Pancreatic cancer is characterized by extremely high mortality and poor prognosis and is projected to be the leading cause of cancer deaths by 2030. Due to the lack of early symptoms and appropriate methods to detect pancreatic carcinoma at an early stage as well as its aggressive progression, the disease is often quite advanced by the time a definite diagnosis is established. The 5-year relative survival rate for all stages is approximately 8%. Therefore, detection of pancreatic cancer at an early surgically resectable stage is the key to decrease mortality and to improve survival. The traditional methods for diagnosing pancreatic cancer involve an imaging test, such as ultrasound or magnetic resonance imaging, paired with a biopsy of the mass in question. These methods are often expensive, time consuming, and require trained professionals to use the instruments and analyze the imaging. To overcome these issues, biosensors have been proposed as a promising tool for the early diagnosis of pancreatic cancer. The present review critically discusses the latest developments in biosensors for the early diagnosis of pancreatic cancer. Protein and microRNA biomarkers of pancreatic cancer and corresponding biosensors for pancreatic cancer diagnosis have been reviewed, and all these cases demonstrate that the emerging biosensors are becoming an increasingly relevant alternative to traditional techniques. In addition, we discuss the existing problems in biosensors and future challenges.

Biocontrol of Salmonella Typhimurium in Raw Salmon Fillets and Scallop Adductors by Using Bacteriophage SLMP1.

The aim of this study was to evaluate the effectiveness of bacteriophage (phage) SLMP1 to reduce Salmonella Typhimurium on contaminated raw salmon fillets and scallop adductors as a function of Salmonella inoculum level, phage dose, storage temperature, and storage time. Samples were inoculated with 102 and 104 CFU/g Salmonella and then treated with different concentrations of phage SLMP1, followed by incubation at 4, 15, and 25°C, respectively. The results showed that 108 PFU/g was the optimal concentration of phage for the control of Salmonella, which was applied in the following storage experiments over a 7-day period at 4°C, a 4-day period at 15°C, and a 2-day period at 25°C. For the salmon fillets samples, 102 CFU/g Salmonella could be reduced below the detection limit at all three temperatures, whereas 104 CFU/g Salmonella was first decreased and then increased at 15 and 25°C. For the scallop adductors samples, 102 CFU/g Salmonella could be reduced below the detection limit first and then increased after a certain period at 15 and 25°C. The variation trends of 104 CFU/g Salmonella in scallop adductors were similar to those in salmon fillets. The results also showed that the Salmonella counts of both inoculum levels on samples could be reduced below the detection limit or maintained at a low level by phage SLMP1 during storage at 4°C. Phage SLMP1 remained stable on raw salmon fillets and scallop adductors. This study indicated that phage SLMP1 has potential effectiveness as a biocontrol agent of Salmonella in seafood.

miR-124 downregulation leads to breast cancer progression via LncRNA-MALAT1 regulation and CDK4/E2F1 signal activation.

The long non-coding RNA (lncRNA) metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) has been recently shown to be dysregulated in several cancers. However, the mechanisms underlying the role of MALAT1 in breast cancer remain unclear. Herein, we showed that MALAT1 was aberrantly increased in breast cancer tissues and cells. MALAT1-siRNA inhibited breast cancer cell proliferation and cell cycle progression in vitro and in vivo. Furthermore, MALAT1 acted as an endogenous potent regulator by directly binding to miR-124 and down-regulating miR-124 expression. In addition, MALAT1 reversed the inhibitory effect of miR-124 on breast cancer proliferation and was involved in the cyclin-dependent kinase 4 (CDK4) expression. Taken together, our data highlight the pivotal role of MALAT1 in breast cancer tumorigenesis. Moreover, the present study elucidated the MALAT1-miR-124-CDK4/E2F1 signaling pathway in breast cancer, which might provide a new approach for tackling breast cancer.

[ß-Glucan promotes the maturation and migration of bone marrow-derived dendritic cells].

OBJECTIVE: To investigate the effects of ß-glucan on the maturation and migration of bone marrow-derived dendritic cells (BMDCs). METHODS: BMDCs were isolated from mouse bone marrow cells in vitro and induced by ß-glucan for maturation. The expressions of cell surface markers were detected by flow cytometry (FCM). The cytokines (IL-6, IL-12p40, tumor necrosis factor α) in the supernatants were measured by ELISA, and the expressions of intracellular CC chemokine receptor 1 (CCR1), CCR2, CCR5, CCR7 were determined by real-time quantitative PCR. Furthermore, the chemotactic response to CC chemokine ligand 19 (CCLl9) and CCL21, i.e. CCR7-1igands, was measured by Transwell(TM) migration assay. Moreover, the number of migrated cells in the draining lymph nodes was analyzed by FCM. RESULTS: Compared with the control group, the expressions of co-stimulation molecules (MHC II, CD40, CD80, CD86) on BMDCs were up-regulated in the presence of ß-glucan. Furthermore, ß-glucan could prompt BMDCs to secret high levels of IL-6, TNF-α, IL-12 p40 and increase the production of CCR7 mRNA. After ß-glucan treatment, BMDCs were more sensitive to CCL19/CCL21. The number of BMDCs migrated from subcutaneous injection site into the draining lymph nodes significantly increased in ß-glucan group. CONCLUSION: ß-glucan can promote the maturation of BMDCs and enhance the migration ability of BMDCs in vitro and in vivo.

ß-glucan restores tumor-educated dendritic cell maturation to enhance antitumor immune responses.

Tumors can induce the generation and accumulation of immunosuppressive cells such as myeloid-derived suppressor cells (MDSCs) in a tumor microenvironment, contributing to tumor escape from immunological attack. Although dendritic cell-based cancer vaccines can initiate antitumor immune responses, tumor-educated dendritic cells (TEDCs) involved in the tolerance induction have attracted much attention recently. In this study, we investigated the effect of ß-glucan on TEDCs and found that ß-glucan treatment could promote the maturation and migration of TEDCs and that the suppressive function of TEDCs was significantly decreased. Treatment with ß-glucan drastically decreased the levels of regulatory T (Treg) cells but increased the infiltration of macrophages, granulocytes and DCs in tumor masses, thus elicited Th1 differentiation and cytotoxic T-lymphocyte responses and led to a delay in tumor progression. These findings reveal that ß-glucan can inhibit the regulatory function of TEDCs, therefore revealing a novel function for ß-glucan in immunotherapy and suggesting its potential clinical benefit. ß-Glucan directly abrogated tumor-educated dendritic cells-associated immune suppression, promoted Th1 differentiation and cytotoxic T-lymphocyte priming and improved antitumor responses.

MiR-124 inhibits cell proliferation in breast cancer through downregulation of CDK4.

Studies have shown that microRNAs (miRNAs) are involved in the malignant progression of human cancer. However, little is known about the potential role of miRNAs in breast carcinogenesis. miR-124 expression in breast cancer tissue was measured by quantitative real-time PCR (qRT-PCR). Target prediction algorithms and luciferase reporter gene assays were used to investigate the target of miR-124. Breast cancer cells growth was regulated by overexpression or knockdown miR-124. At the end of the study, tumor-bearing mice were tested to confirm the function of miR-124 in breast cancer. In this study, we demonstrated that the expression of miR-124 was significantly downregulated in breast cancer tissues compared with matched adjacent non-neoplastic tissues. We identified and confirmed that cyclin-dependent kinase 4 (CDK4) was a direct target of miR-124. Overexpression of miR-124 suppressed CDK4 protein expression and attenuated cell viability, proliferation, and cell cycle progression in MCF-7 and MDA-MB-435S breast cancer cells in vitro. Overexpression of CDK4 partially rescued the inhibitory effect of miR-124 in the breast cancer cells. Moreover, we found that miR-124 overexpression effectively repressed tumor growth in xenograft animal experiments. Our results demonstrate that miR-124 functions as a growth-suppressive miRNA and plays an important role in inhibiting tumorigenesis by targeting CDK4.

Protective effect of YHI and HHI-I against experimental acute pancreatitis in rabbits.

AIM:To observe the protective effect of combined i.v. administraction of Yuanhu injection (YHI) and Huoxuehuayu injection-I (HHI-I) against acute pancreatitis (AP) in rabbits.METHODS:Sever acute pancreatitis (SAP) was induced by retrograde infusion of artificial bile juice into biliary-pancreatic duct, and treated with YHI and HHI-I intravenously. The protective effect was judged by the survival time and rate, serum amylase, serum interleukin-6, pancreatic microcirculation and pathological alteration.RESULTS:Combined use of YHI and HHI-I could markedly increase the rabbits' 5-day survival rate after AP (83.3% in the treatment group and 33.3% in control). The serum amylase value (x-± s) decreased to 1596.6U/L± 760.50U/L in the 5th day from the high level (6320.83U/L± 2614.12U/L) in the 1st day after AP in the treatment group, while in the control group the amylase activity in the 5th day was 2095.0U/L± 1081.87U/L, being significantly different from that before AP (837.17U/L± 189.12U/L). YHI and HHI-I also obviously improved the pancreatic microcirculation and lowered the serum interleukin-6 level, one of the indices of severe pancreatitis. Pathological examination indicated all the changes typical for AP in YHI and HHI-I treatment group were milder than those in the control.CONCLUSION:YHI and HHI-I used in combination might have protective effect against acute pancreatitis in rabbits.