Inhibition of microRNA-448 suppresses CD4+ T cell inflammatory activation via up-regulating suppressor of cytokine signaling 5 in systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease. MicroRNA-448 (miR-448) has a pro-inflammatory effect in various inflammation-related diseases and is up-regulated in serum of patients with SLE. However, the role of miR-448 in SLE development remains elusive. In our study, we found high expression of miR-448 in peripheral blood mononuclear cells (PBMCs) of SLE patients, and miR-448 level was positively associated with disease severity. Besides, miR-448 level was up-regulated during the growth of MRL/lpr mice. To investigate the function of miR-448 in SLE, we subjected 8-week MRL/lpr mice to injection of lentivirus (LV)-mediated anti-miR-448. Inhibition of miR-448 reduced serum IgG and anti-dsDNA IgG contents, 24 h urine protein and blood urea nitrogen (BUN) levels, increased complement C3 concentration, and ameliorated splenomegaly and lymphadenectasis in MRL/lpr mice. MiR-448 inhibition alleviated renal inflammatory infiltration and glycogen deposition. Moreover, miR-448 inhibition promoted Treg cell activation and inhibited Th17 cell proportion in naïve CD4+ T cells from spleens, along with elevated interleukin (IL)-10 and reduced IL-17A levels. In vitro, miR-448 inhibition diminished CD4+ T cell polarization toward Th17 cells under Th17-polarizing conditions. Further, luciferase reporter assay revealed that miR-448 binds to the 3'UTR of suppressor of cytokine signaling 5 (SOCS5). SOCS5 expression was down-regulated in the spleens of MRL/lpr mice and induced Th17 cells. SOCS5 deficiency partially reversed the role of miR-448 in Th17 differentiation and IL-17A expression in SLE. Taken together, inhibition of miR-448 impedes Th17 cell activation and tissue damages via targeting SOCS5 in SLE.

Histone methyltransferase EZH2 in proliferation, invasion, and migration of fibroblast-like synoviocytes in rheumatoid arthritis.

OBJECTIVE: Rheumatoid arthritis (RA) may lead to irreversible joint damage. The role of histone modifications in RA has been emphasized. This study investigated the effect of histone methyltransferase EZH2 on fibroblast-like synoviocytes (FLSs) in RA. MATERIALS AND METHODS: Synovial tissues were collected from RA patients and non-RA patients (NC). RA-FLSs and NC-FLSs were isolated and identified using flow cytometry. EZH2 expression in synovial tissues and FLSs was detected using RT-qPCR and Western blot. The proliferation, migration, and invasion of RA-FLSs and NC-FLSs were measured using MTT, EdU, and Transwell assays. The binding of EZH2, H3K27me3, and miR-22-3p was analyzed using ChIP assay. The targeting relationship between miR-22-3p and CYR61 was verified using dual-luciferase assay. miR-22-3p and CYR61 expressions were detected using RT-qPCR. CYR61 and H3K27me3 levels were detected using Western blot. Functional rescue experiments were performed to verify the effect of miR-22-3p or CYR61 on RA-FLSs. RESULTS: EZH2 was highly expressed in synovial tissues and FLSs from RA patients. The proliferation, migration, and invasion ability of RA-FLSs was stronger than that of NC-FLSs. Downregulation of EZH2 repressed proliferation, migration, and invasion of RA-FLSs. EZH2 inhibited miR-22-3p expression by binding to the miR-22-3p promoter and increasing H3K27me3 methylation level, and thereby upregulated CYR61 expression. Downregulation of miR-22-3p or overexpression of CYR61 annulled the inhibitory effect of EZH2 silencing on RA-FLS proliferation, migration, and invasion. CONCLUSION: EZH2 bound to the miR-22-3p promoter and inhibited miR-22-3p expression by upregulating H3K27me3 level, thereby promoting CYR61 expression and inducing the proliferation, migration, and invasion of RA-FLSs.

Disabled-2 overexpression mediates immune suppression in systemic lupus erythematosus by modulating Treg/Th17 cell differentiation.

Systemic lupus erythematosus (SLE) is an autoimmune disorder. T helper 17 (Th17) and regulatory T (Treg) cells play key roles in SLE progression. Disabled-2 (DAB2) exhibits immunomodulatory effects in inflammatory diseases. However, the role of DAB2 in SLE and the precise mechanisms remain unknown. Here, a decreased DAB2 expression and an increased miR-448-3p level were observed in peripheral blood mononuclear cells (PBMCs) from SLE patients. DAB2 level was negatively correlated with SLE Disease Activity Index (SLEDAI), suggesting a functional correlation between DAB2 and SLE. To test this, we used 8-week-old MRL/lpr mice and treated them with lentivirus-mediated DAB2 (LV-DAB2) or its negative control (LV-NC). LV-DAB2 treatment increased DAB2 expression and reduced serum immunoglobulin G (IgG) and anti-dsDNA IgG levels. DAB2 upregulation alleviated splenomegaly and lymphadenopathy and SLE-related organ damage. Moreover, DAB2 enhanced the percentage of CD25+ Foxp3+ Treg cells, but reduced Th17 cell frequency in lupus, along with the reduction in tumour necrosis factor α (TNF-α), interleukin (IL)-6 (IL-6) and IL-17A levels, and the elevation in IL-10. In vitro, naive CD4+ T cells isolated from MRL/lpr mice were polarized into Th17 or Treg phenotypes and treated with lentivirus. LV-DAB2 treatment downregulated IL-17A expression and inhibited the generation of CD4+ IL-17A+ Th17 cells. DAB2 triggered the production of IL-10 and the activation of Treg cells. Furthermore, DAB2 was verified as a direct target for miR-448-3p. MiR-448-3p overexpression cancelled the promoting effect of DAB2 on Treg cell differentiation. Taken together, DAB2 exerts an immunosuppressive effect on SLE through promoting Treg cell activation and inhibiting Th17 cell differentiation, which may be modulated by miR-448-3p.

Quercetin ameliorates salivary gland apoptosis and inflammation in primary Sjögren's syndrome through regulation of the leptin/OB-R signaling.

Dry mouth is the main manifestation of Sjögren syndrome (SS). Quercetin has been reported to alleviate radiation-induced salivary gland damage, yet the effect of quercetin on SS-caused salivary gland damage remains unclear. This study aimed to investigate the effects of quercetin on SS-induced salivary gland damage and the mechanism underlying its therapeutic potential in SS. Here, NOD/Ltj mice were used to spontaneously mimic SS-induced salivary gland inflammation in vivo and salivary gland epithelial cells (SGECs) were stimulated by interferon-γ (IFN-γ) to mimic cell inflammation in vitro. Results showed that quercetin significantly reduced loss of saliva flow, salivary gland damage, cell apoptosis, and inflammatory response in NOD/Ltj mice. Quercetin treatment also significantly reduced the increased serum leptin (LP) levels in NOD/Ltj mice. Furthermore, quercetin blocked the increases in the expression of obesity receptor (OB-R) and its downstream Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling in the salivary glands. In vitro experiments confirmed that quercetin could protect SGECs from IFN-γ-induced cell apoptosis and inflammation through the LP/OB-R-activated JAK2/STAT3 signaling. Hence, quercetin might protect against SS-induced salivary gland damage by relieving cell apoptosis and inflammation by inhibiting the LP/OB-R signaling, providing a new perspective for treating SS-induced dry mouth.

Analysis of red blood cells' dynamic status in a simulated blood circulation system using an ultrahigh-speed simultaneous framing optical electronic camera.

Alterations in the morphologic and mechanical properties of red blood cells (RBCs) are considered direct indicators of blood quality. Current measures of characterizing these properties in vivo are limited by the complicated hemodynamic environment. To better evaluate the quality of fresh and stored blood, a new research platform was constructed to evaluate the hemodynamic characteristics of RBCs. The research platform consists mostly of a microfluidic chip, microscope, and ultrahigh-speed simultaneous framing optical electronic camera (USFOEC). The microfluidic chip was designed to simplify the complicated hemodynamic environment. The RBCs were diluted in erythrocyte preservative fluid and infused into the microfluidic channels. After approximately 600× magnification of using the microscope and camera, the RBCs' dynamic images were captured by the USFOEC. Eight sequential and blur-free images were simultaneously captured by the USFOEC system. Results showed that RBC deformation changed with flow velocity and stored RBCs were less sensitive to deformation (Kfresh < Kstored ). The frozen-stored RBCs were better able to sustain hydrodynamic stress (DI49day = 0.128 vs. DIfrozen = 0.118) than cold-stored RBCs but more sensitive to variations in flow speed (K49day = 1626.2 vs. Kfrozen = 1318.2). Results showed that the stored RBCs had worse deformability than fresh RBCs, but frozen-stored RBCs may incur less damage during storage than those stored at merely cold temperatures. This USFOEC imaging system can serve as a platform for direct observation of cell morphological and mechanical properties in a medium similar to a physiologic environment. © 2016 International Society for Advancement of Cytometry.

Oxymatrine induces apoptosis in human cervical cancer cells through guanine nucleotide depletion.

Oxymatrine is an alkaloid obtained primarily from Sophora roots and has been shown to show anticancer effects in various cancers. However, the cellular and molecular effects of this agent on cervical cancer have been poorly characterized. Here, we investigated the antitumor effect of oxymatrine on a human cervical cancer cell line (HeLa). Our results showed that application of oxymatrine significantly inhibited the cell growth and tumorigenesis in a dose-dependent manner and induced apoptosis through caspase-dependent pathways as determined using flow cytometry and TUNEL staining analysis. To define the proteins potentially related to the mechanisms of action, proteomic analysis was utilized to detect proteins altered by oxymatrine. As the downregulated gene, inosine monophosphate dehydrogenase type II (IMPDH2) was responsible for oxymatrine-induced mitochondrial-related apoptosis. Moreover, oxymatrine depleted intracellular guanosine 5'-triphosphate (GTP) levels by effective IMPDH inhibition. Functional analyses further showed that oxymatrine and tiazofurin, an inhibitor of IMPDH2, sensitized resistant HeLa/DDP cells to cisplatin. In addition, the expression of IMPDH2 in cervical cancer was significantly higher than that in the normal cervical epithelium. Taken together, these findings suggest that targeting of IMPDH2 by potential pharmacological inhibitors, oxymatrine in combination with chemotherapy, might be a promising means of overcoming chemoresistance in cervical cancer with high IMPDH2 expression, and may thus provide new insights into the mechanism of oxyamtrine-induced anticancer effects.

Visual analysis of the research frontiers, hotspots and trends of exercise therapy intervention in tumor-related sleep-wake disorders.

Objective: To systematically understand the research frontiers, hotspots and development trends of exercise therapy in the intervention of tumor-related sleep-wake disorders, and to provide scientific basis for follow-up research. Methods: Downloaded the original research papers on February 26, 2024, from the Web of Science core collection database, on tumor-associated sleep-wake disorders. The data that met the inclusion criteria were imported into the Bibliometric Analysis Platform (http://biblimetric.com), CiteSpace 6.3.R1 and VOSviwer1.6.20 software for visual analysis, and imported into Excel2021. Scientometric analysis was performed with Oringin2021 and PyCharm Community Edition 2022.1.3. Results: A total of 512 original research papers on tumor-related sleep-wake disorders were obtained. The most influential countries in the subject area are the United States, Spain and German, the institutions are the University of California System, Sun Yat Sen University and Northwestern University, et al., the authors are Berger AM, Aaronson NK, Bower JE, et al., and the journals are Cancer, Brit J Cancer and Cancer Nurs. The co-cited references suggest that the current research frontier in the field mainly involves the level, place and program of exercise therapy, including the relationship between physical activity, sedentary behavior and cancer prevention and control. The results of co-occurrence keyword network analysis showed that quality of life, physical activity, breast cancer, exercise, fatigue, and survivors may be the research hotspots in this field, with breast cancer, health, aerobic exercise, adults, and chemotherapy being the most popular. Conclusions: The number of papers published and the research enthusiasm in this field show a steady upward trend. However, there is a lack of influential institutions and scholars, and there is relatively little research collaboration across countries/regions/institutions. The scientific research influence of institutions and scholars in most European and American countries/regions is significantly ahead of that of institutions and scholars in Asian and African countries/regions. But Sun Yat Sen University in China is a relatively active and influential scientific research institution in recent years, which is worthy of attention. In addition, the research frontier of this discipline is the level, place and program of exercise therapy auxiliary intervention, and the research hotspots involve breast cancer, health, aerobic exercise, adults, chemotherapy, et al. Their clinical efficacy needs to be further demonstrated in multi-center, large-sample and high-quality prospective studies.

Long noncoding RNA H19 synergizes with STAT1 to regulate SNX10 in rheumatoid arthritis.

Erosive destruction of joint structures is an important event in the rheumatoid arthritis (RA) development where fibroblast-like synoviocytes (FLS) represent the main effectors. The implication of long noncoding RNAs (lncRNAs) in RA has not been clearly established. Here, we sought to assess the function of lncRNA H19 in RA by assessing its contribution to the phenotype of FLS. H19 was overexpressed in RA-FLS, and H19 promoted RA-FLS proliferation, invasion as well as angiogenesis and reduced RA-FLS apoptosis. Moreover, H19 loss significantly alleviated joint redness and swelling and reduced inflammatory response, synovial hyperplasia and cartilage damage in arthritic mice induced by collagen. Mechanistically, H19 significantly increased the transcription of sorting nexin (SNX) 10 in RA-FLS by promoting STAT1 translocation into the nucleus. Overexpression of SNX10 or STAT1 mitigated the repressing effects of H19 loss on RA in mice. Our findings highlight that H19 upregulation may result in the development of FLS-mediated RA via the STAT1/SNX10 axis. H19 might serve as a possible therapeutic target for RA treatment.

MicroRNA-142-5p promotes the proliferation and metastasis of nasopharyngeal carcinoma.

Growing pieces of evidence reported abnormal expression of microRNA in various cancer. Our research aimed to ascertain the miR-142-5p expression and its potential function in the growth and metastasis of human nasopharyngeal carcinoma (NPC). In human NPC tissues and cell lines, miR-142-5p expression was quantified via the real-time qPCR assay. Functionally, the potential effect of miR-142-5p in human CNE-1 and SUNE-1 cells through MTT assay, colony formation assay, Transwell assay, and cell cycle assay. In addition, the potential target gene of miR-142-5p was determined by the dual-luciferase reporter assay. MiR-142-5p expression was remarkably elevated in human NPC tissues, CNE-1 and SUNE-1 cells. MiR-142-5p overexpression obviously enhanced the ability of cell proliferative and colony formation, and prevented G1 phase arrest in CNE-1 and SUNE-1 cells. Further, the migration number of NPC cells was increased compared to NP69 cells. BTG3 was identified as the direct target gene of miR-142-5p. Inhibition of BTG3 expression could reverse the cell proliferation by miR-142-5p-induced. Overall, miR-142-5p could strengthen the NPC cell's proliferation and migration by directly targeting BTG3. Hence, miR-142-5p may provide a new strategy and program for future clinical treatment of NPC.

LncRNA FOXD3-AS1 promotes breast cancer progression by mediating ARF6.

BACKGROUND: Breast cancer is one of the most common malignant tumor in women. The high metastatic characteristics cause a high mortality rate of breast cancer. Increasing number of studies have indicated that long non-coding RNAs (lncRNAs) play key roles in the progression of human cancers including breast cancer. In this study, we studied the expression and molecular mechanisms of lncRNA FOXD3-AS1 in breast cancer. METHODS: The expression of lncRNA FOXD3-AS1 was analyzed by TCGA database and RT-qPCR assay. CCK8 assay was used to measure cell proliferation ability. Cell migration and invasion capacities were detected by transwell assay. Potential targets of lncRNA and miRNA were predicted by bioinformatic tools. The targeting relationship between genes was verified by dual-luciferase reporter assay. The nude mice tumor model was performed to study the effect of FOXD3-AS1 on breast cancer in vivo. Protein expression was detected by western blot. RESULTS: In the present study, we found that the FOXD3-AS1 expression was significantly increased in breast cancer tissues compared with normal tissues and involved in the poor prognosis of patients. Functionally, knockdown of FOXD3-AS1 suppressed cell proliferation and metastasis abilities in vitro, and tumor growth in vivo. Mechanistically, FOXD3-AS1 functioned as a competing endogenous RNA (ceRNA) to upregulate ARF6 expression by targeting miR-127-3p. In addition, the roles of FOXD3-AS1 on cell proliferation and metastasis were achieved through miR-127-3p/ARF6 axis. CONCLUSION: In summary, our results reported the regulatory mechanism of FOXD3-AS1 in breast cancer progression by targeting miR-127-3p/ARF6 axis to affect cell proliferation, migration, invasion and tumor growth.

Matrine exerts antitumor activity in cervical cancer by protective autophagy via the Akt/mTOR pathway in vitro and in vivo.

Matrine is a quinazoline alkaloid extracted from Sophora flavescens. The aim of the present study was to determine whether matrine can induce autophagy in the human HeLa and SiHa cervical cancer cell lines in vitro and in vivo. Cell viability assay was used to assess the suppressive effect of matrine and cisplatin on the proliferation of HeLa and SiHa cells. A total of 28 4-week-old female BALB/c nude mice were used for the in vivo study. Autophagy and protein expression were observed via transmission electron microscopy, monodansylcadaverine and immunohistochemical staining and western blotting. The inhibitory effect of matrine on the proliferation of cervical cancer cells was time- and dose-dependent. The combination of matrine and cisplatin synergistically inhibited the proliferation of cervical cancer cells in vitro and in vivo. Transmission electron microscopy showed that after the addition of matrine, numerous autophagosomes and autophagolysosomes were observable in HeLa and SiHa cells, as demonstrated by monodansylcadaverine staining. Western blotting and immunohistochemical staining showed that as the concentration of matrine increased, the expression of the autophagy marker LC3A/B-II also increased significantly in vitro and in vivo. These findings suggested that matrine inhibited the proliferation of cervical cancer cells and induced autophagy by inhibiting the Akt/mTOR signaling pathway. Thus, matrine may represented a potential candidate in combination therapy for cervical cancer as an inducer of autophagy.

Mesenchymal Stem Cell-Originated Exosomal Circular RNA circFBXW7 Attenuates Cell Proliferation, Migration and Inflammation of Fibroblast-Like Synoviocytes by Targeting miR-216a-3p/HDAC4 in Rheumatoid Arthritis.

BACKGROUND: Rheumatoid arthritis (RA) is a chronic autoimmune disease of articular joint damage and elevated synovial hyperplasia. Abnormal proliferation, invasion inflammatory response of rheumatoid fibroblast-like synoviocytes (RA-FLS) play a critical role in RA progression. Mesenchymal stem cell (MSC)-derived exosomal circular RNAs are promising therapeutic manner for disease treatment. This work aimed to decipher the role of exosomal circFBXW7 in RA. METHODS: The expression of circFBXW7, miR-216a-3p, and HDAC4 were detected in clinical RA samples. The RA rat model was established. Isolation and identification of exosomes from MSCs was conducted. The effects of exosomal circFBXW7 on RA was evaluated by qPCR, CCK-8, transwell assays, flow cytometry, Western blotting, ELISA, and immunohistochemical assay. Interaction between miR-216a-3p and circFBXW7 or HDAC4 was determined by luciferase reporter gene assay and RNA pulldown. RESULTS: Exosomal circFBXW7 treatment suppressed proliferation, migration and inflammatory response of RA-FLSs and damage of RA model. CircFBXW7 could directly sponge miR-216a-3p to upregulate the expression of HDAC4. Inhibition of HDAC4 or upregulation of miR-216a-3p abolished the therapeutic function of exosomal circFBXW7. Our data demonstrated that circFBXW7 and HDAC4 were decreased, and miR-216a-3p was elevated in clinical RA sample compared with healthy samples. CONCLUSION: We concluded that MSC-derived exosomal circFBXW7 suppressed proliferation, migration and inflammatory response of RA-FLSs and damage of RA rats via sponging miR-216a-3p and release the activation of HDAC4. These findings may provide a novel therapeutic target for RA.

The clinical effect of aspirin combined with low-molecular-weight heparin in the treatment of severe preeclampsia and the combination's effect on pregnancy outcomes.

OBJECTIVE: To explore the clinical effects of aspirin combined with low-molecular-weight heparin (LMWH) in the treatment of patients with severe preeclampsia and the combination's influence on pregnancy outcomes. METHODS: From October 2018 to June 2020, 104 patients with severe preeclampsia who underwent treatment in our hospital were recruited as the study cohort and divided into two groups according to different treatment scheme each patient underwent. In the research group (RG), the 54 patients were administered aspirin combined with LMWH, and the other 50 patients in the control group (CG) were administered routine treatment. The total effective rates were compared between the two groups. The blood pressure, coagulation function, hemorheology, and renal function indexes were compared before and after the therapy. The Apgar scores of the newborns and the incidences of adverse pregnancy outcomes were measured at 1 and 5 minutes after the births. RESULTS: After the therapy, the systolic blood pressure (SBP) and the diastolic blood pressure (DBP) in the RG were lower than they were in the CG. The PT and APTT in the RG were significantly higher than they were in the CG, and the FIB and D-D were significantly lower than they were in the CG. After the treatment, the hematocrit, the erythrocyte sedimentation rate, and the plasma viscosity in the RG were significantly lower than they were in the CG. The 24 h UP, BUN, UA, and Scr levels in the RG were significantly lower than they were in the CG. The Apgar scores of the newborns in the RG were significantly higher than they were in the CG at 1 min and 5 min after the births. After the therapy, the incidence of adverse pregnancy outcomes in the RG was significantly lower than it was in the CG, and the total effective rate in the RG was significantly higher than it was in the CG. CONCLUSION: Aspirin combined with LMWH can effectively improve the clinical efficacy, the coagulation function, the renal function, and the blood pressure levels, and the combination can reduce adverse pregnancy outcomes in severe preeclampsia patients.

miR-99a and -99b inhibit cervical cancer cell proliferation and invasion by targeting mTOR signaling pathway.

MicroRNAs were demonstrated to play an important role in the regulation of gene expression. Here, we showed that miR-99a and -99b (miR-99a/b) were down-regulated in human cervical cancer patient tissues and were negatively related with lymphatic metastasis. In addition, overexpression of miR-99a/b inhibited cell growth and invasion, whereas suppression of miR-99a/b yielded the reverse phenotype. Dual luciferase report assay revealed that mTOR was identified as a novel target gene of both miR-99a and -99b. Altogether, these results suggested that miR-99a/b directly and negatively regulated mTOR expression in cervical cancer cells, and enforced the importance of miR-99a/b and their targets in the malignant phenotypes of cervical carcinogenesis.

[Curcumin inhibits HeLa cell invasion and migration by decreasing inducible nitric oxide synthase].

OBJECTIVE: To investigate the inhibitory effects of curcumin against HeLa cell invasion and migration and explore the underlying mechanisms. METHODS: HeLa cells were exposed to curcumin treatment at the concentrations of 0, 10, 25, 50, 100, 150 and 200 µmol/L for 24 h. MTT and TUNEL assays were used to assess the cell proliferation inhibition and apoptosis, respectively. Transwell assay was used to evaluate the invasiveness and migration of the treated cells, and RT-PCR and Western blotting were employed to detect the changes in the expression of inducible nitric oxide synthase (iNOS), and MMP-9 and E-cad, the 2 markers of cell invasion and migration, were detected by Western blotting. The capacity of NO production in HeLa cells was measured by Griess method. RESULTS: Curcumin inhibited the proliferation of HeLa cells by inducing cell apoptosis in a concentration-dependent manner. Curcumin inhibited the invasion and migration of HeLa cells by increasing E-cad expression and decreasing MMP-9 expression, and also decreased the expression level of iNOS and NO production in the cells. CONCLUSION: Curcumin inhibits the invasion and migration of HeLa cells by decreasing the expression of iNOS.