INTRODUCTION:Esophageal squamous cell carcinoma (ESCC) lacks effective early diagnostic markers. High-dimensional WGCNA (hdWGCNA) enables the systematic identification of macrophage-related signatures from single-cell data, offering a novel approach to discovering potential biomarkers for ESCC. METHODS:This study analyzed bulk transcriptomic datasets and a single‑cell RNA‑seq dataset from the Gene Expression Omnibus (GEO) database. Specifically, the GSE53625 dataset (179 ESCC and 179 matched normal samples) was a training cohort, while GSE20347 (17 ESCC and 17 normal samples) was an independent validation set. The single‑cell dataset GSE196756 (3 ESCC and 3 adjacent normal tissues) was processed to construct a cellular atlas. The CellChat package was used to assess the communication between macrophages and other cell subpopulations. Macrophage‑associated co‑expression modules were identified by hdWGCNA. Differentially expressed genes (DEGs) were screened with the limma package, and the intersection of DEGs and hub genes from hdWGCNA was further refined via LASSO and SVM-RFE algorithms to obtain candidate genes. Functional enrichment, immune infiltration, drug prediction, and molecular docking were subsequently performed. In vitro experiments were carried out to validate the expression and functional role of candidate genes in ESCC cell lines. RESULTS:Four macrophage‑related gene models consisting of TGFBI, FTL, GPNMB, and APOE were created and validated. These genes were significantly overexpressed in ESCC tissues and exhibited high diagnostic accuracy, reaching an AUC value >0.8 in both the training and independent validation cohorts. In vitro functional assays confirmed that silencing TGFBI suppressed ESCC cell proliferation, migration, and invasion. Enrichment analysis linked these genes to neutrophil activation and apoptosis pathways, and immune infiltration analysis further revealed their correlation with immunosuppressive cell subsets. Drug prediction yielded 190 candidate compounds, and molecular docking suggested potential binding of retinoic acid, rosiglitazone, and GDC‑0941 to the corresponding targets in ESCC. DISCUSSION:In this study, four characteristic genes (TGFBI, FTL, GPNMB, and APOE) were identified for the diagnosis of ESCC, contributing to the improvement of early detection of the cancer. CONCLUSION:The present findings contributed to the pathogenesis research, detection, and management of ESCC.

The features of high heterogeneity and aggressiveness of glioblastoma (GBM) predispose it to temozolomide (TMZ) resistance, which notably impairs therapeutic efficacy. Cytokine family members have been shown to activate pro-survival pathways in GBM cells and facilitate the formation of immunosuppressive microenvironments, contributing to TMZ resistance in GBM. The present study aimed to investigate the role of cytokine IL-37 in the malignant progression and TMZ resistance of GBM. Recombinant human IL-37b (rIL-37) enhanced the survival of TMZ-treated GBM U251 cells. IL-37 silencing or overexpression notably reduced or increased the survival rate of U251 cells exposed to TMZ, respectively. In TMZ-resistant U251 (U251-TR) cells, IL-37 knockdown also improved cellular sensitivity to TMZ. Consistent with these findings, rIL-37 treatment promoted proliferation and suppressed apoptosis of U251 cells under TMZ exposure, whereas IL-37 silencing significantly suppressed growth and enhanced the apoptotic responses of U251-TR cells receiving TMZ. To explore the underlying mechanisms, RNA sequencing was performed and revealed that the U251 cells undergoing TMZ exposure and rIL-37 treatment exhibited an altered transcriptomic profile, characterized by the activation of both mitogen-activated protein kinase (MAPK) pathways and senescence-related pathways. After confirming the activation of cellular MAPK cascades and senescence, Adezmapimod, a p38 MAPK-specific inhibitor, effectively counteracted rIL-37-induced improvements in cellular survival and proliferative capacity, suppression of apoptosis and activation of senescence in U251 cells under TMZ exposure, indicating the necessity of the MAPK pathway in TMZ resistance. The present study expands the current knowledge of the roles of IL-37 during the malignant progression of GBM and demonstrates that IL-37 enhances TMZ resistance in GBM cells by activating the MAPK pathway and inhibiting apoptosis, suggesting that targeted inhibition of MAPK cascades in GBM is promising in future clinical practice.

Abstract 18β-Glycyrrhetinic acid (GA) exhibits excellent anti-inflammatory and antioxidant activities, and has attracted considerable attention in the field of cerebral ischemia−reperfusion (I/R) injury therapy. However, it is plagued by problems of low bioavailability and dose-dependent toxicity. As a drug delivery platform for the treatment of ischemic stroke, microenvironment-responsive liposomes can cross the impaired blood−brain barrier, efficiently deliver drugs to the lesion sites, and improve therapeutic efficacy. Herein, to address the challenges posed by the highly oxidative-stress microenvironment of I/R and the intrinsic physicochemical properties of GA, we encapsulated GA into liposomes and incorporated thioketal (TK) linkages into the phospholipid bilayer to fabricate GA@TK-Lip. GA@TK-Lip can effectively target damaged neurons, attenuate neuroinflammation, scavenge excessive ROS, ameliorate cellular oxidative stress via the Nrf2/HO-1 pathway, reduce neuronal apoptosis and cerebral infarct volume to 43.2% and 23.3% of that in the MCAO group, and restore neurological function. Collectively, this study provides novel insights into the development of promising therapeutic strategies for ischemic stroke.

This study aimed to identify and validate aging-related genes (ARGs) implicated in multiple myeloma (MM), thereby advancing the understanding of the molecular mechanisms underlying the disease. mRNA expression data were retrieved from the Gene Expression Omnibus, and used as a training set to identify ARGs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were conducted to explore the functional roles of the ARGs in MM. Candidate genes identified through least absolute shrinkage and selection operator (LASSO) logistic regression and support vector machine recursive feature elimination (SVM-RFE) were compared using a Venn diagram, which revealed the overlap between the genes identified by the two algorithms. A receiver operating characteristic curve was generated based on the screening results. The candidate genes were further validated using the GSE39754 and GSE5900 datasets. Real-time quantitative polymerase chain reaction (RT-qPCR) was employed to validate the mRNA expression of key genes. Compared to normal individuals, patients with MM exhibited differential expression of 19 genes, of which 5 were upregulated and 14 downregulated. A total of 6 candidate genes (TXN, JUN, FOS, HIF1A, CAT and KCNA3) were identified through LASSO regression and SVM-RFE screening. Among them, TXN exhibited the most significant differential expression, suggesting its potential as a diagnostic biomarker for MM. In addition, in vitro RT-qPCR analysis confirmed that the mRNA levels of TXN in MM cells aligned with the bioinformatics findings, showing higher expression compared to normal B-lymphocyte cell lines. In conclusion, this study identified age-associated molecular patterns in MM and highlighted the diagnostic potential of TXN, offering novel insights for clinical applications in MM.

Tobacco smoking and alcohol consumption are significant risk factors for esophageal squamous cell carcinoma (ESCC), with their Collaborative interaction amplifying carcinogenic risks. To elucidate the role and underlying mechanisms of tobacco-alcohol co-exposure in ESCC pathogenesis, we treated the immortalized human normal esophageal epithelial cell line NE-1 and C57BL/6 mice with 4-nitroquinoline-1-oxide (4NQO, a tobacco carcinogen analog) combined with ethanol. Results demonstrated that 4NQO-ethanol co-exposure accelerated esophageal squamous carcinogenesis. Through network toxicology and in vitro/in vivo experiments, we further validated that 4NQO-ethanol Combine exacerbated inflammatory responses and aggravated DNA damage. Mechanistically, 4NQO-ethanol co-exposure activated the TNF-α/TRAF2/NF-κB signaling pathway. Furthermore, the addition of the TNF-α inhibitor reduced the proliferative and invasive capacities of cells co-stimulated by 4NQO and ethanol. These findings indicate that tobacco-ethanol co-exposure promotes the development of esophageal squamous cell carcinoma by inducing inflammation and DNA damage through TNF-α/TRAF2/NF-κB signaling.

Copper is an important trace element required by mammals and is widely involved in regulating various life activities. Overload or imbalance of copper ions can activate specific signaling pathways, leading to impaired oocyte function, decreased ovulation efficiency, and impaired embryo implantation. However, whether these changes are related to cuproptosis, a copper-driven, mitochondrial form of regulated cell death triggered by proteotoxic stress, a newly discovered phenomenon in recent years, remains unclear. In this study, combination treatment with a specific concentration of CuSO4 and elesclomol, a copper ionophore, delivered Cu+/Cu2+ to the mitochondrial matrix and potentiated cuproptosis, which triggered cuproptosis, leading to a decreased ability of cumulus cells to synthesize progesterone and estrogen. Moreover, during the maturation of oocytes, the expansion of cumulus cells and the levels of electron transport chain-related proteins decreased, whereas the calcium ion level, CaMKIIγ protein level, and phosphorylation level of CX43Ser255 significantly increased. In addition, although cuproptosis in cumulus cells did not directly induce cuproptosis in oocytes, it decreased the quality and further maturation of oocytes. During this process, abnormalities in mitochondrial distribution and function, cortical granule distribution, spindle morphology, and intracellular cAMP levels were detected in oocytes. The protein level of CDK1 decreased, whereas that of Emi1 increased. These results suggest that in cumulus cells, cuproptosis impairs mitochondrial function and triggers abnormal intracellular calcium homeostasis, which subsequently leads to overactivation of CaMKIIγ, blockage of Cx43 channels between COCs, and abnormal accumulation of cAMP in oocytes. Consequently, aberrant expression of the cell cycle regulatory factors CDK1 and Emi1 ultimately inhibits the in vitro maturation of oocytes. These findings clarify the effect of cuproptosis on bovine oocyte maturation, which contributes to a deeper understanding of the regulatory mechanisms governing oocyte maturation and may provide new directions for optimizing in vitro oocyte culture systems.

Background:Gestational diabetes mellitus (GDM), a prevalent metabolic complication during pregnancy, has a global prevalence of approximately 14%. Its onset is closely associated with insulin resistance, insufficient compensatory function of β - cells, and abnormal placental function. Epidemiological studies have indicated that type 2 diabetes is an independent risk factor for breast cancer. However, the association between GDM and the risk of breast cancer remains controversial. Objective:This systematic review and meta-analysis aim to comprehensively evaluate the association between GDM and the risk of breast cancer and explore its underlying mechanisms. Methods:This study systematically searched PubMed, Web of Science, Scopus, EMBASE, and the Cochrane Library databases, covering the period from establishing each database until April 14, 2025. Two researchers extracted relevant data and assessed the quality of included studies using the Newcastle-Ottawa Scale. The study evaluated inter-study heterogeneity using the I² statistic. Based on the magnitude of heterogeneity, fixed-effect or random-effect models were employed to calculate the pooled hazard ratio (HR) and its corresponding 95% confidence interval (CI). Additionally, subgroup analyses, sensitivity analyses, funnel plot analyses, and publication bias assessments were performed. All data analyses were conducted using STATA 17 software. Results:The overall analysis revealed no significant association between GDM and breast cancer risk (HR=1.03, 95%CI: 0.92-1.15). However, subgroup analysis revealed significant regional heterogeneity: within the regional subgroups, North American results showed an association between GDM and a reduced breast cancer risk (HR=0.89, 95%CI: 0.84-0.95), whereas Asian findings suggested an association with an increased risk (HR=1.23, 95%CI: 1.15-1.31). No significant associations were observed in subgroups based on study design (cohort/case-control) or follow-up duration (short-term/long-term). Sensitivity analysis demonstrated robust results, and there was no publication bias in this study. Conclusion:In summary, there is no significant association between GDM and breast cancer risk overall. However, notable regional heterogeneity exists: in the North American subgroup, GDM is associated with a reduced risk of breast cancer, while in the Asian subgroup, GDM is significantly associated with an increased risk of breast cancer. Systematic Review Registration:https://www.crd.york.ac.uk/PROSPERO/, identifier CRD420251032589.

Bone health management for breast cancer spans the entire cycle of patient care, including the prevention and treatment of bone loss caused by early breast cancer treatment, the adjuvant application of bone-modifying agents to improve prognosis, and the diagnosis and treatment of advanced bone metastases. Making good bone health management means formulating appropriate treatment strategies and dealing with adverse drug reactions, and will help to improve patients' quality of life and survival rates. The Breast Cancer Expert Committee of the National Cancer Center for Quality Control organized relevant experts to conduct an in-depth discussion on the full-cycle management of breast cancer bone health based on evidence-based medicine, and put forward reasonable suggestions to guide clinicians to better deal with health issues in bone health clinics.

Endocrine treatment, particularly tamoxifen, has shown significant improvement in the prognosis of patients with estrogen receptor-positive (ER-positive) breast cancer. However, the clinical utility of this treatment is often hindered by the development of endocrine resistance. Therefore, a comprehensive understanding of the underlying mechanisms driving ER-positive breast cancer carcinogenesis and endocrine resistance is crucial to overcome this clinical challenge. In this study, we investigated the expression of MICAL-L2 in ER-positive breast cancer and its impact on patient prognosis. We observed a significant upregulation of MICAL-L2 expression in ER-positive breast cancer, which correlated with a poorer prognosis in these patients. Furthermore, we found that estrogen-ERβ signaling promoted the expression of MICAL-L2. Functionally, our study demonstrated that MICAL-L2 not only played an oncogenic role in ER-positive breast cancer tumorigenesis but also influenced the sensitivity of ER-positive breast cancer cells to tamoxifen. Mechanistically, as an estrogen-responsive gene, MICAL-L2 facilitated the activation of the Akt/mTOR signaling pathway in ER-positive breast cancer cells. Collectively, our findings suggest that MICAL-L2 could serve as a potential prognostic marker for ER-positive breast cancer and represent a promising molecular target for improving endocrine treatment and developing therapeutic approaches for this subtype of breast cancer.

IL-10 is a pleiotropic cytokine that plays a significant role in antiviral and antitumor immunity. Potent CD8+ T cells express IL-10 after stimulation by strong TCR signaling, which promotes the killing effect of CD8+ T cells. However, the regulation of IL-10 expression in CD8+ T cells and its signaling pathway to enhance CD8+ T cell function are largely unknown. In this study, we investigated the JAK-STAT signaling molecules that regulate IL-10 expression in CD8+ T cells and the JAK-STAT signaling pathway that IL-10 enhances the function of CD8+ T cells through its receptor, using small molecule inhibitors and CRISPR-Cas9 gene editing. Our findings provide new insights and a theoretical basis for the immunotherapy of tumors.

Myeloid-derived suppressor cells (MDSCs) play a crucial role in maintaining maternal-fetal tolerance by expressing some immune-suppressive molecules, such as indoleamine 2,3-dioxygenase (IDO). Toxoplasma gondii (T. gondii) infection can break the immune microenvironment of maternal-fetal interface, resulting in adverse pregnancy outcomes. However, whether T. gondii affects IDO expression in dMDSCs and the molecular mechanism of its effect are still unclear. Here we show, the mRNA level of IDO is increased but the protein level decreased in infected dMDSCs. Mechanistically, the upregulation of transcriptional levels of IDO in dMDSCs is regulated through STAT3/p52-RelB pathway and the decrease of IDO expression is due to its degradation caused by increased SOCS3 after T. gondii infection. In vivo, the adverse pregnancy outcomes of IDO−/− infected mice are more severe than those of wide-type infected mice and obviously improved after exogenous kynurenine treatment. Also, the reduction of IDO in dMDSCs induced by T. gondii infection results in the downregulation of TGF-β and IL-10 expression in dNK cells regulated through Kyn/AhR/SP1 signal pathway, eventually leading to the dysfunction of dNK cells and contributing the occurrence of adverse pregnancy outcomes. This study reveals a novel molecular mechanism in adverse pregnancy outcome induced by T. gondii infection.

Esophageal squamous cell carcinoma (ESCC) is a malignant tumor of the digestive tract with strong migratory and invasive abilities. Gas6 is closely associated with the progression of many malignant tumors; however, the role of Gas6 in the progression of esophageal cancer is unclear. Here, we report that the knockdown of Gas6 inhibited esophageal cancer cell proliferation, migration, and invasion. In addition, Gas6 knockdown downregulated the levels of P-PI3K and P-AKT. Taken together, the findings confirm that Gas6 knockdown can inhibit esophageal cancer progression and can exert anti-tumor effects on esophageal cancer through the PI3K/AKT pathway.

Cancer-associated fibroblasts (CAFs) are an important part of tumour microenvironment, but its role in immunotherapy of gastric cancer (GC) is still needed to further study. In this study, we firstly distinguish the GC related CAFs via single cell sequencing dataset. CAFs in deep layers of GC tissues gain more developmental potential. Moreover, we found Glypican-3 (GPC3) is up-regulated in the CAFs subgroups of the advanced GC and correlated with poor prognosis in GC patients. In addition, higher GPC3 expression GC patients have higher TIDE (Tumour Immune Dysfunction and Exclusion) score, dysfunction and exclusion score. independent GC cohort also show GC patients with GPC3high CAFs have lower response rate to PD-1 therapy. GPC3 secreted from CAFs up-regulated PD-L1, TIM3, CD24, CYCLIN D1, cMYC and PDK mRNA expression level in HGC-27 cells. At last, in vivo model demonstrate that targeting GPC3high CAFs sensitizing the PD-1 blockage therapy in GC. In conclusion, GPC3 expression in CAFs is a critical prognostic biomarker, and targeting GPC3high cancer-associated fibroblasts sensitizing the PD-1 blockage therapy in GC.Key messagesGlypican-3 (GPC3) is up-regulated in the CAFs subgroups of the advanced gastric cancer.Gastric cancer patients with GPC3high CAFs have lower response rate to PD-1 therapy.Targeting GPC3high CAFs sensitizing the PD-1 blockage therapy in gastric cancer.

Objective. This study is aimed at investigating the role of lncRNA GHET1 in the progression of triple-negative breast cancer (TNBC). Methods. Tumor tissues and paracancerous tissues (normal) of TNBC patients were collected. Human normal breast cells (MCF10A) and TNBC cells (MDA-MB-468 and HCC1937) were employed for in vitro analysis. The expression of lncRNA GHET1, miR-377-3p, and GRSF1 was detected by qRT-PCR. The lncRNA GHET1 and miR-377-3p were overexpressed or knocked down in the TNBC cells, respectively. To determine the specific biological activities of the TNBC cells, MTT, flow cytometry, and wound healing assay were adopted to evaluate the cellular proliferation, apoptosis, and migration abilities, respectively. MMP-9 and MMP-2 protein expression levels were detected as well by Western blot in the cells. The relationship between miR-377-3p and lncRNA GHET1, miR-377-3p, and GRSF1 was validated using dual-luciferase reporter assay. Results. lncRNA GHET1 was significantly upregulated in the TNBC patients' tissues and the TNBC cell lines. Overexpression of lncRNA GHET1 significantly increased the proliferation and migration ability, but decreased apoptosis in the TNBC cells. Additionally, overexpression of lncRNA GHET1 upregulated both MMP-9 and MMP-2 protein expression levels. Correlation analysis found that miR-377-3p had a positive relationship with GRSF1, but had a negative relationship with lncRNA GHET1. miR-377-3p mimic attenuated the effects of lncRNA GHET1 on cellular proliferation, apoptosis, and migration of the TNBC cells. Conclusion. lncRNA GHET1 promotes TNBC progression through the miR-377-3p/GRSF1 signaling axis.

MICAL1 has been reported to be involved in the malignant processes of several types of cancer cells, however, the roles of MICAL1 in colorectal cancer (CRC) have not been well-characterized. This study aims to investigate the cellular functions and molecular mechanisms of MICAL1 in CRC cells. Here, we found that both mRNA and protein levels of MICAL1 were down-regulated in colorectal cancer tissues compared with matched adjacent non-tumor tissues, and the expression level of MICAL1 was correlated with the metastatic status of colorectal cancer. Importantly, overexpression of MICAL1 significantly inhibited colorectal cancer cell migration and growth, and increased the level of E-cadherin and Occludin, and suppressed the expression level of Vimentin and N-cadherin; while silencing of MICAL1 promoted CRC cell migration and enhanced EMT. In addition, MICAL1 overexpression significantly inhibited the proliferation and growth of CRC in vitro and in vivo. Moreover, RNA sequencing and bioinformatics analysis identified that MICAL1 was closely correlated with "cell migration", "cell cycle" and "β-catenin signaling" genesets. Mechanistically, overexpression of MICAL1 downregulated the mRNA level of EGR1 and β-catenin, decreased the protein level and nuclear translocation of β-catenin, and inhibited the transcriptions of β-catenin downstream targets, c-myc and cyclin D1. The ectopic expression of EGR1 or β-catenin can significantly block the MICAL1-mediated inhibitory effects. Collectively, MICAL1 is down-regulated in CRC, and plays an inhibitory role in the migration and growth of CRC cells by suppressing the ERG1/β-catenin signaling pathway.

Abstract Background Toxoplasma gondii infection during pregnancy can lead to fetal defect(s) or congenital complications. The inhibitory molecule B7-H4 expressed on decidual macrophages (dMφ) plays an important role in maternal–fetal tolerance. However, the effect of B7-H4 on the function of dMφ during T. gondii infection remains unclear. Methods Changes in B7-H4 expression on dMφ after T. gondii infection were explored both in vivo and in vitro. B7-H4-/- pregnant mice (pregnant mice with B7-H4 gene knockout) and purified primary human dMφ treated with B7-H4 neutralizing antibody were used to explore the role of B7-H4 signaling on regulating the membrane molecules, synthesis of arginine metabolic enzymes and cytokine production by dMφ with T. gondii infection. Also, adoptive transfer of dMφ from wild-type (WT) pregnant mice or B7-H4-/- pregnant mice to infected B7-H4-/- pregnant mice was used to examine the effect of B7-H4 on adverse pregnancy outcomes induced by T. gondii infection. Results The results illustrated that B7-H4-/- pregnant mice infected by T. gondii had poorer pregnancy outcomes than their wild-type counterparts. The expression of B7-H4 on dMφ significantly decreased after T. gondii infection, which resulted in the polarization of dMφ from the M2 toward the M1 phenotype by changing the expression of membrane molecules (CD80, CD86, CD163, CD206), synthesis of arginine metabolic enzymes (Arg-1, iNOS) and production of cytokines (IL-10, TNF-α) production. Also, we found that the B7-H4 downregulation after T. gondii infection increased iNOS and TNF-α expression mediated through the JAK2/STAT1 signaling pathway. In addition, adoptive transfer of dMφ from a WT pregnant mouse donor rather than from a B7-H4-/- pregnant mouse donor was able to improve adverse pregnancy outcomes induced by T. gondii infection. Conclusions The results demonstrated that the downregulation of B7-H4 induced by T. gondii infection led to the dysfunction of decidual macrophages and contributed to abnormal pregnancy outcomes. Moreover, adoptive transfer of B7-H4+ dMφ could improve adverse pregnancy outcomes induced by T. gondii infection. Graphical Abstract

Molecule interacting with CasL-like protein 2 (MICALL2) is believed to regulate cytoskeleton dynamics, tight junction formation, and neurite outgrowth. However, its biological role and the underlying mechanism in colorectal cancer (CRC) remain largely elusive. qRT-PCR, Western blotting and immunohistochemistry assays were used to detect the expression levels of different genes. Next, mass spectrometry, co-immunoprecipitation and immunofluorescence staining were used to detect the interactions of proteins. Furthermore, MTT assay, colony formation assay, wound-healing assays and xenograft tumor models were performed to demonstrate the functions of MICALL2 in CRC. In addition, transcriptome sequencing and Western blotting were conducted to verify the mechanism of MICALL2 in CRC. We found that both mRNA and protein levels of MICALL2 are up-regulated in colorectal cancer tissues compared with non-tumor tissues and that its overexpression is closely correlated with poor prognosis. Ubiquitin E3 ligase Tripartite motif-containing protein 21 (TRIM21) mediated MICALL2 ubiquitination and proteasome-dependent degradation, negatively correlated with MICALL2 levels, and reversely regulated the tumorigenic activity of MICALL2 in CRC. Functional studies confirmed that MICALL2 promoted colorectal cancer cell growth and migration via the Wnt/β-catenin signaling pathway. As a substrate of ubiquitinase TRIM21, MICALL2 enhances the growth and migration of colorectal cancer cells and activates the Wnt/β-catenin signaling pathway.

Objective . This study is aimed at investigating the role of lncRNA GHET1 in the progression of triple-negative breast cancer (TNBC). Methods . Tumor tissues and paracancerous tissues (normal) of TNBC patients were collected. Human normal breast cells (MCF10A) and TNBC cells (MDA-MB-468 and HCC1937) were employed for in vitro analysis. The expression of lncRNA GHET1, miR-377-3p, and GRSF1 was detected by qRT-PCR. The lncRNA GHET1 and miR-377-3p were overexpressed or knocked down in the TNBC cells, respectively. To determine the specific biological activities of the TNBC cells, MTT, flow cytometry, and wound healing assay were adopted to evaluate the cellular proliferation, apoptosis, and migration abilities, respectively. MMP-9 and MMP-2 protein expression levels were detected as well by Western blot in the cells. The relationship between miR-377-3p and lncRNA GHET1, miR-377-3p, and GRSF1 was validated using dual-luciferase reporter assay. Results . lncRNA GHET1 was significantly upregulated in the TNBC patients’ tissues and the TNBC cell lines. Overexpression of lncRNA GHET1 significantly increased the proliferation and migration ability, but decreased apoptosis in the TNBC cells. Additionally, overexpression of lncRNA GHET1 upregulated both MMP-9 and MMP-2 protein expression levels. Correlation analysis found that miR-377-3p had a positive relationship with GRSF1, but had a negative relationship with lncRNA GHET1. miR-377-3p mimic attenuated the effects of lncRNA GHET1 on cellular proliferation, apoptosis, and migration of the TNBC cells. Conclusion . lncRNA GHET1 promotes TNBC progression through the miR-377-3p/GRSF1 signaling axis.

BackgroundLung squamous cell carcinoma (LUSC) is a major subtype of non-small cell lung cancer. The tumor immune microenvironment (TIME) affects the anti-tumor immune response and the patient’s prognosis, although the TIME in LUSC patients is incompletely understood.MethodsWe retrospectively collected surgical specimens from patients with previously untreated primary LUSC. Histopathological examination was used to identify tumor regions and adjacent regions, and imaging mass cytometry was used to characterize the immune cells in those regions. The results were compared between regions and between patients.ResultsWe identified heterogeneity in the TIME on comparing different patients with LUSC, although the tumor region and adjacent region both exhibited an immune response to the tumor. The TIME typically included a large number of infiltrating and activated T-cells, especially CD8+ T-cells, which closely interacted with the tumor cells in the tumor region. There was limited infiltration of B-cells, NK cells, and NKT cells, while the major immune suppressor cells were CD33+ myeloid-derived cells. We also identified a novel population of CD3−CD4+ cells with high expression of Foxp3 and TNFα, which might modulate the tumor microenvironment and play a proinflammatory role in the TIME.ConclusionsThe TIME of LUSC appears to be immunogenic and heterogenous, with predominant infiltration of activated CD8+ T-cells. The interactions between the tumor cells and T-cells facilitate the anti-tumor activity. A novel subpopulation of CD3−CD4+ cells with high TNFα and Foxp3 expression may modulate the tumor microenvironment and play a proinflammatory role.

In sheep industry, hypothermia caused by insufficient brown adipose tissue (BAT) deposits is one of the major causes of lamb deaths. Enhancing the formation and function of BAT in neonatal lamb increases thermogenesis and hence reduces economic losses. The aim of the present study was to explore the effect and mechanism of melatonin on sheep brown adipocyte formation and function. Sheep brown adipocyte precursor cells (SBACs) isolated from perirenal BAT were treated with melatonin (1 and 10 nM). The SBACs subjected to melatonin exhibited a decreased proliferation ability, accompanied by down-regulated proliferating cell nuclear antigen, cyclin D1, and CDK4 protein contents in a melatonin dose-dependent manner. Melatonin promoted brown adipocyte formation and induced the expression of brown adipogenic markers, including uncoupling protein 1 and PR domain-containing 16 during differentiation of SBAC. Moreover, the AMP-activated protein kinase α1 (AMPKα1) activity was positively correlated with brown adipocyte formation potential. Importantly, melatonin effectively activated AMPKα1. Furthermore, promotional effects of melatonin were abolished by AMPKα1 knockout, suggesting the involvement of AMPKα1 in this process. Collectively, these results suggested that melatonin enhanced brown adipocyte formation in SBACs in vitro through activation of AMPKα1.