Objective:To investigate protective effect and mechanism of Tim-3 on sepsis-induced acute lung injury(ALI)by pro-moting mitophagy of alveolar macrophages and inhibiting activation of NLRP3 inflammasome.Methods:LPS-stimulated mouse alveo-lar macrophage(MH-S)model and sepsis-induced ALI mouse model were constructed.Tim-3 siRNA interference technique was used to knock down Tim-3 expression in MH-S cells,and anti-Tim-3 antibody mice were injected intraperitoneally to block Tim-3 function.Western blot was used to detect protein expressions of NLRP3,ASC,cleaved-caspase-1 and mitophagy-related proteins(LC3B,P62,PINK1 and Parkin)in MH-S cells and lung tissue of mice with sepsis-induced ALI.Laser confocal fluorescence staining was used to measure ROS level and mitochondrial membrane potential of MH-S cells.Pathological examination of lung tissue was performed in mice with sepsis-induced ALI in each group,and degree of lung tissue injury was evaluated by Smith scoring system.Bronchoalveolar lavage fluid(BALF)and lung tissue were collected from mice with ALI induced by sepsis in each group.BCA protein quantification method was used to determine protein concentration in BALF.MPO activity in lung tissue was detected by colorimetry.MDA content in lung tissue was detected by TBA method.LC3B protein expression in lung tissue was detected by immunohistochemistry.Results:In mouse alveolar macrophages,Tim-3 knockdown could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins,increase ROS release,inhibit PINK1/Parkin pathway activation and LC3B protein expression,and reduce mitochondrial membrane potential.In mice with sepsis-induced ALI,Tim-3 functional blockade could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins in lung tissue,aggravate lung pathological injury and pulmonary edema,increase MPO activity and MDA content in lung tissue,and reduce positive rate of LC3B protein.Conclusion:Tim-3 plays a protective role in sepsis-induced ALI by promoting mitophagy in alveolar macrophages and inhibiting NLRP3 inflammasome activation via PINK1/Parkin.

AIM:To investigate the impact of mitochondrial carrier homologous protein 2(MTCH2)on the metastasis of oral squamous cell carcinoma and to elucidate its underlying mechanisms.METHODS:Human tongue squamous cell carcinoma CAL-27 cells were selected as the study model.An MTCH2 inhibition and overexpression model was established using small interfering RNA(siRNA)technology and overexpression plasmids.Based on the experimental design,cells were categorized into the following groups:siRNA negative control(siRNA-NC)group,MTCH2 siRNA(siRNA-MTCH2)group,empty vector(OE-NC)group,and MTCH2 overexpression(OE-MTCH2)group.Transwell as-says and wound healing assays were conducted to assess the invasion and migration capabilities of cells in each group.The morphological changes and distribution of actin microfilaments were examined using a red fluorescent probe.The mito-chondrial membrane potential was evaluated using a mitochondrial membrane potential detection kit.The levels of reactive oxygen species(ROS)were measured using a ROS detection kit.Adenosine triphosphate(ATP)production levels were assessed with an ATP detection kit.The protein expression levels of MTCH2 and matrix metalloproteinase 14(MMP14)were analyzed through cell fluorescence and Western blot techniques.RESULTS:The number of invasive cells in the siRNA-MTCH2 group was significantly reduced,and the cell migration rate was notably decreased.Conversely,the OE-MTCH2 group exhibited a significant increase in both the number of invasive cells and the cell migration rate(P<0.01).The fluorescence intensity of F-actin and MTCH2 in the siRNA-MTCH2 group was significantly diminished,while these in-tensities were markedly elevated in the OE-MTCH2 group(P<0.01).The ROS levels in the siRNA-MTCH2 group were significantly higher than those in the siRNA-NC group,whereas the levels of ATP and mitochondrial membrane potential were significantly lower.Conversely,the ROS levels in the OE-MTCH2 group were significantly lower than those in the OE-NC group,with corresponding increases in ATP levels and mitochondrial membrane potential(P<0.01).The expres-sion levels of MTCH2 and MMP14 proteins in the siRNA-MTCH2 group were significantly lower than those in the siRNA-NC group,while the OE-MTCH2 group showed significantly higher expression levels compared to the OE-NC group(P<0.01).CONCLUSION:The MTCH2 is associated with the development of oral squamous cell carcinoma and may facili-tate the metastasis of CAL-27 cells by regulating mitochondrial function and MMP14 protein expression.Therefore,MTCH2 may represent a novel therapeutic target for the treatment of oral cancer.

AIM:To investigate the impact of mitochondrial carrier homologous protein 2(MTCH2)on the metastasis of oral squamous cell carcinoma and to elucidate its underlying mechanisms.METHODS:Human tongue squamous cell carcinoma CAL-27 cells were selected as the study model.An MTCH2 inhibition and overexpression model was established using small interfering RNA(siRNA)technology and overexpression plasmids.Based on the experimental design,cells were categorized into the following groups:siRNA negative control(siRNA-NC)group,MTCH2 siRNA(siRNA-MTCH2)group,empty vector(OE-NC)group,and MTCH2 overexpression(OE-MTCH2)group.Transwell as-says and wound healing assays were conducted to assess the invasion and migration capabilities of cells in each group.The morphological changes and distribution of actin microfilaments were examined using a red fluorescent probe.The mito-chondrial membrane potential was evaluated using a mitochondrial membrane potential detection kit.The levels of reactive oxygen species(ROS)were measured using a ROS detection kit.Adenosine triphosphate(ATP)production levels were assessed with an ATP detection kit.The protein expression levels of MTCH2 and matrix metalloproteinase 14(MMP14)were analyzed through cell fluorescence and Western blot techniques.RESULTS:The number of invasive cells in the siRNA-MTCH2 group was significantly reduced,and the cell migration rate was notably decreased.Conversely,the OE-MTCH2 group exhibited a significant increase in both the number of invasive cells and the cell migration rate(P<0.01).The fluorescence intensity of F-actin and MTCH2 in the siRNA-MTCH2 group was significantly diminished,while these in-tensities were markedly elevated in the OE-MTCH2 group(P<0.01).The ROS levels in the siRNA-MTCH2 group were significantly higher than those in the siRNA-NC group,whereas the levels of ATP and mitochondrial membrane potential were significantly lower.Conversely,the ROS levels in the OE-MTCH2 group were significantly lower than those in the OE-NC group,with corresponding increases in ATP levels and mitochondrial membrane potential(P<0.01).The expres-sion levels of MTCH2 and MMP14 proteins in the siRNA-MTCH2 group were significantly lower than those in the siRNA-NC group,while the OE-MTCH2 group showed significantly higher expression levels compared to the OE-NC group(P<0.01).CONCLUSION:The MTCH2 is associated with the development of oral squamous cell carcinoma and may facili-tate the metastasis of CAL-27 cells by regulating mitochondrial function and MMP14 protein expression.Therefore,MTCH2 may represent a novel therapeutic target for the treatment of oral cancer.

Objective:To investigate protective effect and mechanism of Tim-3 on sepsis-induced acute lung injury(ALI)by pro-moting mitophagy of alveolar macrophages and inhibiting activation of NLRP3 inflammasome.Methods:LPS-stimulated mouse alveo-lar macrophage(MH-S)model and sepsis-induced ALI mouse model were constructed.Tim-3 siRNA interference technique was used to knock down Tim-3 expression in MH-S cells,and anti-Tim-3 antibody mice were injected intraperitoneally to block Tim-3 function.Western blot was used to detect protein expressions of NLRP3,ASC,cleaved-caspase-1 and mitophagy-related proteins(LC3B,P62,PINK1 and Parkin)in MH-S cells and lung tissue of mice with sepsis-induced ALI.Laser confocal fluorescence staining was used to measure ROS level and mitochondrial membrane potential of MH-S cells.Pathological examination of lung tissue was performed in mice with sepsis-induced ALI in each group,and degree of lung tissue injury was evaluated by Smith scoring system.Bronchoalveolar lavage fluid(BALF)and lung tissue were collected from mice with ALI induced by sepsis in each group.BCA protein quantification method was used to determine protein concentration in BALF.MPO activity in lung tissue was detected by colorimetry.MDA content in lung tissue was detected by TBA method.LC3B protein expression in lung tissue was detected by immunohistochemistry.Results:In mouse alveolar macrophages,Tim-3 knockdown could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins,increase ROS release,inhibit PINK1/Parkin pathway activation and LC3B protein expression,and reduce mitochondrial membrane potential.In mice with sepsis-induced ALI,Tim-3 functional blockade could promote expressions of NLRP3,ASC,cleaved-caspase-1 and P62 proteins in lung tissue,aggravate lung pathological injury and pulmonary edema,increase MPO activity and MDA content in lung tissue,and reduce positive rate of LC3B protein.Conclusion:Tim-3 plays a protective role in sepsis-induced ALI by promoting mitophagy in alveolar macrophages and inhibiting NLRP3 inflammasome activation via PINK1/Parkin.

Epigenomic imbalance drives abnormal transcriptional processes,promoting the onset and progression of cancer.Although defective gene regulation generally affects carcinogenesis and tumor suppression networks,tumor immunogenicity and immune cells involved in antitumor responses may also be affected by epigenomic changes,which may have significant implications for the development and application of epigenetic therapy,cancer immunotherapy,and their combinations.Herein,we focus on the impact of epigenetic regulation on tumor immune cell function and the role of key abnormal epigenetic processes,DNA methylation,histone post-translational modification,and chromatin structure in tumor immunogenicity,and introduce these epigenetic research methods.We emphasize the value of small-molecule inhibitors of epigenetic modulators in enhancing antitumor immune responses and discuss the challenges of developing treatment plans that combine epigenetic therapy and immuno-therapy through the complex interaction between cancer epigenetics and cancer immunology.

Abstract: The differential expression and subcellular localization of single-cell proteins are closely related to the physiological state and pathological mechanisms of the body. The development of single-cell protein in situ imaging methods provides powerful tools for spatial single-cell proteomics research and single-cell protein profiling. This article summarizes the single-cell protein imaging methods developed in recent years, including the circulating immunofluorescence imaging methods based on ordered multi-round antibody incubation, mass spectrometry imaging based on metal element labeled antibodies, fluorescence imaging based on DNA-barcoded antibody, gene encoded fluorescence protein imaging and spectral imaging based on Raman spectroscopy or X-ray spectroscopy, with brief explanation of the imaging principles of these methods. It focuses on the multiple performance, imaging resolution and signal amplification performance of these methods, and analyzes their application characteristics in practical scientific research and clinical work, in the hope of providing some reference for the development of more revolutionary single-cell imaging methods, and promoting the development of biomedical and precision medicine.

OBJECTIVES: Primary tumor treatment through surgical resection and adjuvant therapy has been extensively studied, but there is a lack of effective strategies and drugs for the treatment of tumor metastases. Here, we describe a functional product based on a combination of compounds, which can be used as an adjuvant therapy and has well-known mechanisms for inhibiting cancer metastases, improving anti-cancer treatment, and enhancing immunity and antioxidant capacity. Our designed combination, named MVBL, consists of four inexpensive compounds: L-selenium-methylselenocysteine (MSC), D-‍α‍-tocopheryl succinic acid (VES), β‍-carotene (β‍-Ca), and L-lysine (Lys). METHODS: The effects of MVBL on cell viability, cell cycle, cell apoptosis, cell migration, cell invasion, reactive oxygen species (ROS), and paclitaxel (PTX)-combined treatment were studied in vitro. The inhibition of tumor metastasis, antioxidation, and immune enhancement capacity of MVBL were determined in vivo. RESULTS: MVBL exhibited higher toxicity to tumor cells than to normal cells. It did not significantly affect the cell cycle of cancer cells, but increased their apoptosis. Wound healing, adhesion, and transwell assays showed that MVBL significantly inhibited tumor cell migration, adhesion, and invasion. MVBL sensitized MDA-MB-231 breast cancer cells to PTX, indicating that it can be used as an adjuvant to enhance the therapeutic effect of chemotherapy drugs. In mice, experimental data showed that MVBL inhibited tumor metastasis, prolonged their survival time, and enhanced their antioxidant capacity and immune function. CONCLUSIONS This study revealed the roles of MVBL in improving immunity and antioxidation, preventing tumor growth, and inhibiting metastasis in vitro and in vivo. MVBL may be used as an adjuvant drug in cancer therapy for improving the survival and quality of life of cancer patients.
Mice ; Animals ; beta Carotene ; Lysine/pharmacology* ; Antioxidants/pharmacology* ; Quality of Life ; Paclitaxel/pharmacology* ; Apoptosis ; alpha-Tocopherol ; Succinates/pharmacology* ; Cell Line, Tumor ; Cell Proliferation ; Neoplasms

Atrial fibrillation (AF) is one of the most common arrhythmias, associated with high morbidity, mortality, and healthcare costs, and it places a significant burden on both individuals and society. Anti-arrhythmic drugs are the most commonly used strategy for treating AF. However, drug therapy faces challenges because of its limited efficacy and potential side effects. Catheter ablation is widely used as an alternative treatment for AF. Nevertheless, because the mechanism of AF is not fully understood, the recurrence rate after ablation remains high. In addition, the outcomes of ablation can vary significantly between medical institutions and patients, especially for persistent AF. Therefore, the issue of which ablation strategy is optimal is still far from settled. Computational modeling has the advantages of repeatable operation, low cost, freedom from risk, and complete control, and is a useful tool for not only predicting the results of different ablation strategies on the same model but also finding optimal personalized ablation targets for clinical reference and even guidance. This review summarizes three-dimensional computational modeling simulations of catheter ablation for AF, from the early-stage attempts such as Maze III or circumferential pulmonary vein isolation to the latest advances based on personalized substrate-guided ablation. Finally, we summarize current developments and challenges and provide our perspectives and suggestions for future directions.

To investigate psychological characteristics in different clinical subgroups of insomniacs, and to provide the basis for the accurate simplification of cognitive behavioral therapy for insomnia.
 Methods: A total of 212 insomniacs from November 2014 to June 2017 in Clinical Psychology Department or Sleep Department of 2 general hospitals in Hunan Province were included in convenient and classified into sleep onset insomnia (SOI), difficulty maintaining insomnia (DMI), early morning awakening insomnia (EMAI), and combined insomnia (CI) subgroups. Ford Insomnia Response to Stress Test (FIRST), Simplified Coping Style Questionnaire (SCSQ), Dysfunctional Beliefs and Attitudes about Sleep Scale 16 version (DBAS-16), Sleep-Related Behavior Questionnaire (SRBQ), Pre-sleep Arousal Scale (PSAS), Center for Epidemiological Studies Depression Scale (CES-D), Beck Anxiety Inventory (BAI) were used to investigate the psychological characteristics.
 Results: SOI and CI insomniacs had a higher frequency in use of sleep-related behavior than those with DMI; CI had a higher frequency in use of sleep-related behavior than those with EMAI (all P<0.05). Both SOI and CI insomniacs had a higher level of pre-sleep cognitive arousal than DMI and EMAI (all P<0.05). CI insomniacs noticed more consequences of insomnia and had more worries on insomnia than DMI, and CI insomniacs had more expectations of sleep than SOI (all P<0.05).
 Conclusion: Insomniacs with different clinical subgroups have different features of psychological characteristics. Both the insomnia subgroups and the psychological characteristics should be taken into account when we simplify cognitive behavioral therapy for insomnia (CBT-I) precisely.
Anxiety ; Arousal ; Cognitive Behavioral Therapy ; Humans ; Sleep ; Sleep Initiation and Maintenance Disorders ; Surveys and Questionnaires