Advances in Clinical and Experimental Medicine最新文献

Currently, there is no doubt that hepatitis C virus (HCV) infection is a systemic disease affecting not only the liver but also a range of other organs - extrahepatic manifestations (EHMs). Extrahepatic manifestations usually occur concurrently with liver disease, primarily have an immunological basis, and/or are a consequence of the direct impact of HCV on virtually all organs. The scope of the problem is significant; it has been shown that 30-40% of HCV-infected individuals are affected, which aligns with our own observations. Viral elimination (either spontaneous HCV clearance or as a result of pharmacotherapy) is crucial for the patient's prognosis, both in terms of liver disease and EHM. Achieving a sustained virological response (SVR) only in many cases of EHMs is associated with remission of clinical symptoms of EHMs.

Background: Recurrent miscarriage (RM) affects 1-2% of couples. Maternally expressed gene 3 (MEG3) is aberrantly expressed in RM patients.

Objectives: To investigate a novel regulatory mechanism, we examined the miR-204-5p/Specificity protein 1 (SP1)/DNA methyltransferase 1 (DNMT1)/MEG3 axis in the trophoblast cell line HTR-8/SVneo.

Material and methods: Human trophoblast cell line HTR-8/SVneo was used and cells were transfected with siRNA targeting SP1, miR-204-5p mimics, pcDNA3.1-DNMT1, or their negative controls (NCs). The methylation inhibitor, 5-azadC, was used to treat the cells transfected with pcDNA3.1-SP1. The reverse transcription quantitative polymerase chain reaction (RT-qPCR) method was used to examine the relative RNA levels of SP1, DNMT1 and MEG3. Western blot assay was performed to measure the protein levels of SP1 and DNMT1. The dual-luciferase reporter gene assay was used to validate the miR-204-5p bindings to SP1. Functional assays were utilized to assess cell apoptosis, colony formation, migration, and invasion.

Results: SP1 knockdown inhibited DNMT1 and increased MEG3 expression. The expression of MEG3 was enhanced by methylation inhibition through 5-azadC, but SP1 upregulation reversed this effect. SP1 knockdown increased apoptosis and decreased migration and invasion, which was reversed by DNMT1 overexpression. SP1 was targeted and inhibited by miR-204-5p. miR-204-5p also inhibited DNMT1, and enhanced the expression of MEG3. miR-204-5p inhibited cell proliferation, migration and invasion, and promoted apoptosis. Overexpression of SP1 partially reversed these effects by activating DNMT1 and inhibiting MEG3.

Conclusions: miR-204-5p promoted MEG3 expression in trophoblast cells via SP1-mediated DNMT1 inhibition, leading to reduced cell migration, proliferation and invasion, as well as increased apoptosis. This study reveals a novel regulatory axis in trophoblast cells, providing insights into potential regulatory mechanisms in RM.

Background: Inflammatory response is involved in the pathogenesis of herpes zoster (HZ) and postherpetic neuralgia (PHN).

Objectives: This study aimed to evaluate levels of proinflammatory factors at different stages of HZ and PHN.

Material and methods: A total of 154 patients within 72 h of HZ onset and 30 healthy controls were included. Patients were followed up to 90 days. The levels of interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α) and C-reactive protein (CRP) were measured at baseline and 90 days. The visual analogue scale (VAS) was used to assess the intensity of pain and PHN patients were divided into mild-to-moderate pain and severe pain group.

Results: Interleukin 6, TNF-α and CRP levels in HZ patients at baseline were significantly higher than in healthy controls and decreased as followed up to 90 days. Moreover, PHN patients had a higher level of IL-6, TNF-α or CRP at baseline and 90 days than non-PHN patients. In addition, PHN patients in the severe pain group had a notably higher baseline or 90-day IL-6, TNF-α and CRP level than in the mild-to-moderate pain group. However, the changes of IL-6, TNF-α and CRP levels between 90 days and baseline were significantly less pronounced in the severe pain group than in the mild-to-moderate pain group.

Conclusions: The levels of proinflammatory cytokines were higher in HZ and PHN patients and associated with pain intensity in PNH patients. These findings suggest that repeated measurements of serum proinflammatory cytokines may aid in clinical management and guide anti-inflammatory treatment strategies.

Background: Hepatic stellate cell (HSC) activation is a critical factor in the development of liver fibrosis. Recent research indicates that mesoderm/mesenchyme homeobox 1 (Meox1) contributes to fibrosis in organs like the skin and heart.

Objectives: To investigate the potential impact of Meox1 on HSC activation and provide an available target for hepatic fibrosis research.

Material and methods: The human HSC cell line LX-2 was utilized to investigate the role of Meox1 in HSC activation. Fibrotic gene expression was analyzed, and assays were conducted to assess cell proliferation, migration and the cell cycle.

Results: Meox1 was identified as a positive regulator of HSC activation. We found that transforming growth factor-β1 (TGF-β1) treatment significantly upregulated Meox1 expression in a dose-dependent manner in LX-2 cells, and the expression levels of α-smooth muscle actin (α-SMA), collagen type I (collagen-I) and matrix metalloproteinase-2 (MMP-2) also increased progressively with higher concentrations of TGF-β1. Knockdown of Meox1 via small interfering RNA (siRNA) inhibited TGF-β1-induced expression of HSC activation markers and fibrotic genes, including α-SMA, collagen-I and MMP-2. Conversely, Meox1 overexpression promoted HSC activation, evidenced by increased levels of α-SMA, collagen-I and MMP-2. Meanwhile, Meox1 overexpression accelerated cell proliferation and enhanced cell migration. Additionally, forced expression of Meox1 in LX-2 cells elevated Smad3 phosphorylation level, although TGF-β1 and total Smad3 protein levels remained unchanged. Furthermore, we observed that Meox1 could induce a redistribution of the cell population, extending the G1 phase, and that Meox1-upregulated p21CIP1/WAF1 expression in LX-2 cells was independent of p53.

Conclusions: Our findings suggest that Meox1 plays a pivotal role in HSC activation and may be involved in the canonical TGF-β1/Smad pathway.

Background: Insulin-like growth factor 2 mRNA-binding protein 2 (IGF2BP2) is implicated in various cancers, but its role in modulating ferroptosis and tumor cell behavior in non-small cell lung cancer (NSCLC) remains unclear.

Objectives: This study aimed to investigate how IGF2BP2-mediated N6-methyladenosine (m6A) modification of solute carrier family 7 member 11 (SLC7A11) affects ferroptosis and NSCLC cell viability.

Material and methods: NSCLC H1299 cells were transfected with either IGF2BP2 or SLC7A11 plasmids and corresponding siRNAs. Expression levels of IGF2BP2, SLC7A11 and ferroptosis markers were analyzed using reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. Cell viability was assessed using the Cell Counting Kit-8 (CCK-8) assay. Reactive oxygen species (ROS) and lipid peroxidation levels were measured with flow cytometry and biochemical kits. The RNA immunoprecipitation (RIP) and mRNA stability assays were utilized to explore the interaction between IGF2BP2 and SLC7A11.

Results: IGF2BP2 expression was significantly upregulated in H1299 cells. Overexpression of IGF2BP2 enhanced cell viability and decreased ferroptosis, whereas its knockdown resulted in reduced cell viability and increased ferroptotic activity. IGF2BP2 enhanced SLC7A11 mRNA stability through m6A modification, and SLC7A11 overexpression reversed the effects of IGF2BP2 knockdown. This interaction increased cell viability and reduced ROS and lipid peroxidation.

Conclusions: IGF2BP2 plays a critical role in NSCLC by stabilizing SLC7A11 mRNA via m6A modification, promoting cell proliferation and suppressing ferroptosis. Targeting the IGF2BP2-SLC7A11 axis may offer a promising therapeutic strategy for NSCLC.

Background: Preeclampsia is a serious pregnancy complication with significant maternal and fetal morbidity. Mitophagy plays a crucial role in its pathogenesis. The importance of this study lies in evaluating the role of parkin in preeclampsia, which may offer new insights into the management of this disease.

Objectives: This study was designed to evaluate the role of parkin in preeclampsia.

Material and methods: To induce a preeclampsia model, pregnant female rats were administered N-nitro-L-arginine methyl ester (L-NAME) subcutaneously at a dose of 50 mg/(kg·day) starting on gestational day 14 for 7 consecutive days. Uteroplacental tissues were then collected, and chorionic trophoblast cells were isolated. Systolic blood pressure (SBP) and urine protein content were measured on days 12 and 20 of pregnancy. Hematoxylin-eosin (H&E) staining and TUNEL staining were employed to assess pathological changes and apoptosis in uteroplacental tissues, respectively. Reverse transcription polymerase chain reaction (RT-qPCR) and western blot analysis were performed to evaluate mRNA and protein expression levels associated with cellular function, mitophagy and the PINK1/parkin signaling pathway.

Results: Compared to the negavtive control (NC) group, rats in the model group showed elevated SBP and urine protein levels (p < 0.01). Chorionic trophoblast cells exhibited substantial damage, with significantly increased levels of apoptosis and autophagy. Moreover, parkin mRNA and protein expression levels were markedly upregulated in the model group. Overexpression of parkin in chorionic trophoblast cells enhanced apoptosis and mitophagy, while the autophagy inhibitor 3-methyladenine (3-MA) significantly alleviated the damage caused by overexpression of parkin.

Conclusions: Parkin aggravates the symptoms of preeclampsia by increasing mitophagy and apoptosis.

Background: The coronavirus disease 2019 (COVID-19) pandemic has significantly accelerated the development and use of new healthcare technologies. While younger individuals may have been able to quickly embrace virtual advancements, older adults may still have different needs in terms of health communication.

Objectives: To identify areas of interest and preferred sources of information related to the COVID-19 pandemic among older adults and to verify their eHealth competencies.

Material and methods: The study was conducted between February 2022 and July 2022. It included listeners from the University of the Third Age (U3A) and younger students. Both groups received information about the HealthBuddy+ chatbot, a questionnaire that addressed respondents' interests about COVID-19, and the PL-eHEALS (eHealth Literacy Scale) questionnaire to measure their eHealth competencies.

Results: There were 573 participants in the study (U3A listeners - 303 participants, median age: 73 years (interquartile range (IQR): 69-77); young adult students - 270, median age: 24 years (IQR: 23-24). The primary source of information about COVID-19 for older adults was television (84.5%), and for younger adults, internet (84.4%). Among the older adults, only 17% ever interacted with a chatbot (younger adults - 78% respectively), and 19% considered it a trustworthy source of information on COVID-19 compared to 79% of younger respondents. Older adults and younger adults in our study were most interested in COVID-19 treatment methods (45.5% and 69.3%, respectively), symptoms of the disease (36.6% and 35.2%, respectively) and chronic diseases coexisting with COVID-19 (35.0% and 51.5%, respectively). However, their eHealth competencies were generally low (median (Me): 34; IQR: 30-39) compared to younger adults (Me: 42; IQR: 40-47).

Conclusions: Health education for older adults should be appropriately tailored to their current needs and differentiated. The level of eHealth competencies of older adults suggests that much work remains to narrow the gap between the eHealth competencies of the younger and older generations.

Background: Salvianolic acid F (SAF), an important water-soluble monomeric component, is derived from the herbal medicine Salvia miltiorrhiza (SM) Bunge. Although SAF has been suggested to suppress various cancers, its role in ovarian cancer (OC) and the underlying mechanisms remain largely unclear.

Objectives: This study aimed to investigate the effects of SAF on OC cell growth, invasion, migration, and apoptosis, as well as to elucidate the underlying mechanisms, including those involving the EP300/PI3K/AKT signaling pathway.

Material and methods: In vitro cell culture experiments were conducted to assess the effects of SAF on the proliferation, migration, invasion, and apoptosis of OVCAR-3 (Ovarian Cancer Cell Line 3) and SK-OV-3 (Sloan-Kettering Ovarian Cancer 3) cells. Network pharmacology was further employed to explore SAF's impact on OC and to elucidate the potential underlying mechanisms. The EP300-mediated PI3K/AKT signaling pathway was selected for validation to confirm SAF's effects on inducing apoptosis and inhibiting cell proliferation in these OC cell lines.

Results: Salvianolic acid F suppressed the growth, invasion and migration of SK-OV-3 and OVCAR-3 cells, and induced apoptosis. A network pharmacology analysis of SAF's effects on OC identified core targets, TP53, EP300, STAT3, MMP9, NFKB1, HIF1A, and PTGS2, through protein-protein interaction (PPI) network analysis using the STRING database. Salvianolic acid F inhibited EP300 expression in SK-OV-3 cells, reduced the p-PI3K/PI3K ratio, and increased both the Bax/Bcl-2 ratio and the cleaved caspase-3/caspase-3 ratio in OVCAR-3 and SK-OV-3 cells. However, the addition of A485, an EP300 inhibitor, did not further enhance the effects of SAF.

Conclusions: Salvianolic acid F inhibited OC cell growth, migration and invasion while promoting apoptosis. The EP300/PI3K/AKT pathway is a key mechanism through which SAF regulates OC progression. Additionally, SAF may represent a promising candidate drug for treating OC.

Strengthening primary healthcare (PHC) research is vital to address the demands of a rapidly changing health landscape. Leadership, infrastructure, and sufficient funding have been discussed as key factors in expanding PHC research capacity. This editorial aims to highlight emerging research priorities in a world increasingly affected by crises such as war, conflict, and climate change. Research on suffering, meaning, hope, and compassion represents a promising and necessary new frontier in PHC. This field urgently needs the attention of academic institutions and funding bodies committed to strengthening primary care and family medicine.

Background: Non-alcoholic fatty liver disease (NAFLD) is currently a public health problem that affects many people worldwide. New-generation dietary agents can contribute to disease control. Also, prediction of liver disease activity with serum markers is an important factor to reduce the need for liver biopsies.

Objectives: The aim of this study was to compare the effects of 20 mg/kg green coffee bean extract (GCBE, standardized to 10 mg/kg chlorogenic acid), 10 mg/kg chlorogenic acid (CA) and 50 mg/kg CA on NAFLD activity score (NAS). We also aimed to create a NAS prediction model as an alternative to liver biopsies.

Material and methods: Male Wistar rats were fed either a high-fat high-cholesterol (HFHC) diet (NAFLD groups, n = 32) or a normal diet (control groups, n = 24). Green coffee bean extract (20 mg/kg) and CA (10 mg/kg and 50 mg/kg) were given by daily oral gavage. At the end of the study, blood samples and liver tissues were collected. Biochemical parameters, glyoxalase 1 (GLO1) and oxidative stress parameters were analyzed. An experienced histopathologist graded NAS.

Results: Green coffee bean extract and 50 mg/kg CA significantly reduced NAS (p < 0.01 and p < 0.001, respectively). HFHC-diet induced NAFLD was associated with higher serum GLO1 levels (p < 0.001). Serum malondialdehyde (MDA) levels were positively correlated with NAS while GLO1 was negatively correlated with NAS (p < 0.001, r = 0.698; p < 0.05, r = -0.367, respectively). Serum MDA and triglyceride were found to be statistically significant in predicting NAS (p < 0.001).

Conclusions: Our study suggests that GCBE and CA can both ameliorate the development of NAFLD. Also, low-dose GCBE and high-dose CA showed similar hepatoprotective effects. Increased GLO1 in NAFLD may be a defense mechanism which is enhanced by GCBE and CA. Moreover, serum MDA and triglyceride levels are promising in NAS prediction.