Objective To explore the effects of circ_100284 in affecting the invasion of esophageal squamous cell carcinoma (ESCC) cells and their sensitivity to 5-fluorouracil (5-FU) chemotherapy via regulating miR-217/mitogen-activated protein kinase 1 (MAPK1) acting as a competing endogenous RNA (ceRNA). Methods The bioinformatics approach was used to analyze the differentially expressed circRNAs in ESCC. qRT-PCR was used to detect the expression of circ_100284 in ESCC tissues and cells. The 5-FU-resistant KYSE450 cell line (KYSE45-R) was established by increasing concentrations of 5-FU. The IC₅₀ of 5-FU in KYSE450 and KYSE450-R cells was determined through MTT assay. qRT-PCR was used to detect the expression of circ_100284, miR-217, and MAPK1 mRNA in KYSE450 and KYSE450-R cells, while Western blotting detecting the protein expression of MAPK1. In the experiments with KYSE450-R cells, we set up the following groups: (1) blank group (without treatment), si-NC group (transfected with si-NC), si-circ_100284 group (transfected with si-circ_100284), pc-Control group (transfected with pc-Control), pc-circ_100284 group (transfected with pc-circ_100284), pc-circ_100284+mimic NC group (transfected with pc-circ_100284 and mimic NC), and pc-circ_100284+miR-217 mimic group (transfected with pc-circ_100284 and miR-217 mimic). These groups were subjected to MTT assay to detect cell viability, Transwell assay to detect cell invasion, and flow cytometry to detect cell apoptosis. (2) Blank group (without treatment), si-NC group (transfected with si-MAPK1 negative control), si-MAPK1 group (transfected with si-MAPK1), si-MAPK1+inhibitor NC group (transfected with si-MAPK1 and inhibitor NC), and si-MAPK1+miR-217 inhibitor group (transfected with si-MAPK1 and miR-217 inhibitor). We detected the mRNA and protein expression of MAPK1 using qRT-PCR and Western blotting. We evaluated cell viability using MTT assay, invasion with Transwell assay, and apoptosis by flow cytometry. circ_100284-WT or circ_100284-MUT reporter plasmids, as well as MAPK1-WT or MAPK1-MUT reporter plasmids, were co-transfected with miR-NC or miR-217 mimic into KYSE450-R cells for 48 h, and dual luciferase reporter assay was used to measure luciferase activity. Results The bioinformatics analysis revealed significant upregulation of circ_100284 in ESCC. Compared with adjacent normal tissues, the expression of circ_100284 in ESCC tissues is enhanced (P<0.05); compared with the HECC cells, the TE-11, ECA109 and KYSE450 ESCC cell lines showed enhanced expression of circ_100284 (P<0.05). Compared with the KYSE450 cells, KYSE450-R cells demonstrated increased IC₅₀ with enhanced expression of circ_100284 and MAPK1 but suppressed expression of miR-217 (P<0.05). Compared with the si-NC group, the si-circ_100284 group demonstrated inhibited invasion and proliferation of cells with increased apoptosis (P<0.05). Compared with pc-Control group, the invasion and proliferation of cells in the pc-circ_100284 group are increased, and cell apoptosis is decreased (P<0.05). Over-expression of miR-217 reversed the malignant biological behavior of ESCC cells induced by pc-circ_100284 (P<0.05). Compared with si-NC group, in the si-MAPK1 group, we observed decreased cell invasion and proliferation, and increased apoptosis (P<0.05), but miR-217 inhibitor reversed the effect of si-MAPK1 on the biological behavior of ESCC cells (P<0.05). The targeting relationship of circ_100284 and miR-217, miR-217 and MAPK1 is confirmed. Conclusion circ_100284 promotes ESCC cell invasion by regulating miR-217/MAPK1, inhibits the chemosensitivity of ESCC cells to 5-FU, and acts as a tumor-promoting factor in ESCC.

Objective To investigate the molecular mechanism of verbascoside against acute lung injury (ALI) by network pharmacology and molecular docking methods, and to validate the findings experimentally. Methods The 2D structure of verbascoside was obtained from the Pubchem database. Active ingredient targets of verbascoside were acquired from Pharmmapper database and Swiss Target Prediction database. Active component targets of ALI were acquired from datebase such as Gene Cards, OMIM, and DisGeNET. Common targets between verbascoside and ALI were determined by overlapping these sets. PPI network for potential targets was constructed using String database and Cytoscape software. The intersection targets were imported into the DAVID database for enrichment analysis of GO biological processes, KEGG signaling pathway and the pathway target genes. Molecular docking between verbascoside and core targets was performed using Autodock vina software. The mRNA expression level of core genes was validated using real-time quantitative PCR (RT-qPCR), and the expression of related proteins was detected using Western blotting. Results A total of 150 target genes of verbascoside against ALI were screened, and the key targets of verbascoside against ALI mainly involve pathways such as Rap1 signaling pathway, PI3K-Akt signaling pathway and MAPK signaling pathway. Verbascoside docked well with the core target molecules. RT-qPCR results showed that, compared with the control group, the mRNA expression levels of HSP90AA1, ALB, TP53, TNF, INS, and HRAS were significantly decreased in cells after the effect of verbascoside (P<0.05); Western blotting indicated that, compared with the model group, verbascoside treatment significantly reduced the expression of p-Akt, p-p38, and p-ERK proteins (P<0.05). Conclusion Verbascoside could inhibit MAPK, Rap1 and PI3K/Akt signaling pathways to exert its anti-ALI effects.

Objective To investigate the effect and mechanism of leucine zipper/EF-hand-containing transmembrane protein 1 (LETM1) on proliferation, migration, apoptosis, osteogenic differentiation, and tumorigenesis in vivo of human osteosarcoma MG63 and 143B cells. Methods The osteosarcoma MG63 and 143B cells were divided into blank control group (without adenovirus infection), negative control group (sh-NC group, infected with RNAi negative control virus), and LETM1 knockdown group (sh-LETM1 group, infected with sh-LETM1 adenovirus). Western blotting was performed to detect LETM1 expression in normal human osteoblasts hFOB1.19 and osteosarcoma cells, and to verify the knockdown effect of adenovirus; cell clone formation assays and CCK-8 method were used to detect the proliferation of MG63 and 143B cells; wound-healing assay and Transwell assay were used to test cell migration; DAPI staining and Annexin V-APC/7-AAD flow cytometry double staining were used to detect the apoptosis of MG63 and 143B cells; alkaline phosphatase (ALP) staining and Alizarin Red S staining were used to evaluate early and late osteogenic differentiation of MG63 and 143B cells. Ten nude mice were divided into sh-NC group (n=5, injected subcutaneously into nude mice with 143B cells infected with RNAi negative control virus) and sh-LETM1 group (n=5, injected subcutaneously into nude mice with 143B cells infected with sh-LETM1 adenovirus), and nude mice subcutaneous tumor formation assay was used to examine the in vivo tumor-forming ability of 143B cells in each group. Results Western blotting showed that the expression of LETM1 protein in osteosarcoma MG63 and 143B cells was significantly higher than that in human normal osteoblasts hFOB1.19 (P<0.05), and that the expression of LETM1 protein was markedly reduced after injection with sh-LETM1 adenovirus in MG63 and 143B cells. The results of cell clone formation assay and CCK-8 assay indicated that in MG63 and 143B osteosarcoma cells, the clone formation ability and proliferation ability were significantly reduced in sh-LETM1 group compared with sh-NC group and blank control group (P<0.01). The results of wound-healing assay and Transwell assay demonstrated that in MG63 and 143B osteosarcoma cells, the cell migration rate in sh-LETM1 group was significantly lower than that in sh-NC group and blank control group (P<0.01). DAPI staining and flow cytometry results revealed that the apoptosis rate in sh-LETM1 group was significantly higher than those in MG63 and 143B osteosarcoma cells in sh-NC group (P<0.01). Alkaline phosphatase staining and Alizarin red S staining experiments showed more stained areas and calcium salt nodules in MG63 and 143B osteosarcoma cells in sh-LETM1 group than those in sh-NC group and blank control group. The results of the subcutaneous tumor formation assay in nude mice indicated that subcutaneous tumor formation ability was reduced in 143B sh-LETM1 group compared with 143B sh-NC group. Conclusion LETM1 promotes the proliferation, migration and in vivo tumor formation of MG63 and 143B osteosarcoma cells and the mechanism may be related to the inhibition of apoptosis and osteogenic differentiation.

Remazolam is a γ-aminobutyric acid (GABA) receptor agonist. It is a short-acting benzodiazepine drug that has just been launched in China in recent years. Its mechanism of action is to enhance inhibitory neurotransmitters of GABA in the central nervous system. It has been widely used in clinical practice for sedation, hypnosis, anti anxiety, and the treatment of insomnia. Meanwhile, remazolam is mainly used for induction and maintenance of general anesthesia during the perioperative period, with a fast onset time and a brief duration of action. Recent studies have shown that remazolam can protect important organs and alleviate organ damage by reducing inflammatory responses and oxidative stress, regulating cell apoptosis, and other pathways. This article summarizes the protective effects and mechanisms of remazolam on the brain, heart, liver, and lung, providing a theoretical basis for the clinical application of remazolam.

Objective To report the clinical characteristics, diagnosis, and treatment process of two infants with infantile epileptic spasms syndrome (IESS) caused by DYNC1H1 mutation, and to review the relevant literature. Methods A retrospective analysis was conducted on the clinical data of two IESS patients with DYNC1H1 mutations who were treated at the First Medical Center of Chinese PLA General Hospital. Databases such as PubMed, Online Mendelian Inheritance in Man (OMIM), China National Knowledge Infrastructure (CNKI), and Wanfang Data Knowledge Service Platform were searched to obtain relevant literature, aiming to summarize the clinical characteristics of IESS patients with DYNC1H1 mutations, and to explore the relationship between treatment and phenotype-genotype. Results Two IESS patients with DYNC1H1 mutations were identified (case 1: c.874C>T, p.Arg292Trp; case 2: c.5884C>T, p.Arg1962Cys). Both patients presented with the onset of spastic seizures in infancy, which were poorly controlled with various medications. They exhibited severe developmental delay, and cranial magnetic resonance imaging in case 1 revealed pachygyria. A search of multiple databases and manual screening yielded a total of 7 English articles and 2 Chinese articles. Fifteen cases of DYNC1H1-related IESS were identified, of which 12 cases progressed to drug-resistant epilepsy and 12 cases had significant congenital structural abnormalities of the cranium. Nine different mutation sites were distributed in 3 structural domains, including 4 cases in the tail domain, 3 cases in the motor with ATPase subunit domain, and 2 cases in the stalk or microtubule-binding domain. Conclusions DYNC1H1 gene variations can cause IESS, often accompanied by brain developmental abnormalities and developmental delay/intellectual disability. The poor prognosis may be attributed to the combined effects of gene dysfunction and brain developmental abnormalities.

With the in-depth study of molecular biology, non-small cell lung cancer (NSCLC) has opened the era of precision medicine based on mutation-based molecular targeting therapy. Epidermal growth factor receptor (EGFR) driver mutations are closely related to the progression of NSCLC, and EGFR-tyrosine kinase inhibitors (TKIs) developed based on this have achieved significant therapeutic effects, but acquired drug resistance is still one of the major factors limiting their long-term use. As resistance mechanisms are further investigated, in addition to secondary EGFR mutation, MET amplification, HER2 amplification, histologic transformation, etc., receptor tyrosine kinase (RTK) fusion mutation have been shown to be a targetable mechanism of acquired resistance. Among the acquired RTK fusion mutations, rearranged during transfection (RET) fusion mutations are the accessible targets of our concern. As the RET molecule continues to be explored, drugs targeting RET fusions have been approved and marketed. There are different clinical strategies to deal with acquired RET fusion mutation mediating resistance to EGFR-TKIs treatment. In this review, the structure and function of RET, its relationship with EGFR-TKIs resistance, and treatment strategies are reviewed to further improve patient survival outcomes.

Objective To compare the fidelity of chronic obstructive pulmonary disease (COPD) models established using two methods: exposure to cigarette smoke (CS) and exposure to motor vehicle exhaust (MVE) in rats. Methods Twenty-four male SD rats were randomly divided into control, CS-exposed (CS), and MVE-exposed (MVE) groups, with 8 rats per group. Rats in CS and MVE groups were exposed to CS or MVE, respectively, to induce COPD models. After COPD model established, lung function of each group was assessed. Bronchoalveolar lavage fluid (BALF) was collected to measure inflammatory cell counts, levels of inflammatory cytokines interleukin-6 (IL-6) and tumor necrosis factor (TNF)‑α, and expression levels of mucin 5AC (MUC5AC). Lung tissue sections were stained with hematoxylin and eosin (HE) to observe pulmonary tissue and airway pathological changes. Periodic acid-Schiff (PAS) staining was used to detect goblet cell hyperplasia in airways. Results Compared with control group, rats in CS and MVE groups showed significantly increased inspiratory resistance (RI), total lung capacity (TLC), and lung static compliance (Cchord) (P<0.05), while expiratory flow parameters FEV₅₀/FVC were significantly decreased (P<0.05). Compared with MVE group, rats in CS group had significantly higher R_I, TLC, and Cchord (P<0.05), and lower FEV₅₀/FVC (P<0.05). HE staining of lung tissues showed that mean linear intercept (MLI) was significantly higher in both CS and MVE groups compared with control group (P<0.05), with CS group having higher MLI than MVE group (P<0.05). BALF analysis revealed that white blood cells, neutrophils, macrophages, lymphocytes, IL-6, and TNF-α levels were significantly higher in both CS and MVE groups compared with control group (P<0.05), and inflammatory cell counts, IL-6, and TNF-α levels were higher in CS group compared with MVE group(P<0.05). PAS staining of lung tissues indicated that goblet cells in large airways were significantly increased in both CS and MVE groups compared with control group (P<0.05), with CS group showing higher goblet cell counts than MVE group (P<0.05). Expression levels of MUC5AC in BALF were significantly higher in both CS and MVE groups compared with control group (P<0.05), with CS group having significantly higher MUC5AC levels than MVE group (P<0.05). Conclusions Exposure to CS or MVE can establish a rat model of COPD, with CS exposure better mimicking characteristics of acute exacerbation of COPD compared to MVE exposure.

Objective To analyze the clinical characteristics of high energy metabolism in lung cancer patients and its correlation with body composition, nutritional status, and quality of life, and to develop a corresponding risk prediction model. Methods Retrospectively analyzed 132 primary lung cancer patients admitted to the First Hospital of Shanxi Medical University from January 2022 to May 2023, and categorized into high (n=94) and low energy metabolism group (n=38) based on their metabolic status. Differences in clinical data, body composition, Patient Generated Subjective Global Assessment (PG-SGA) scores, and European Organization for Research and treatment of Cancer (EORTC) Quality of Life Questionnaire-Core 30 (QLQ-C30) scores were compared between the two groups. Logistic regression was used to identify the risk factors for high energy metabolism in lung cancer patients, and a risk prediction model was established accordingly; the Hosmer-Lemeshow test was used to assess the model fit, and the ROC curve was used to test the predictive efficacy of the model. Results Of the 132 patients with primary lung cancer, 94 (71.2%) exhibited high energy metabolism. Compared with low energy metabolism group, patients in high-energy metabolism group had a smoking index of 400 or higher, advanced disease staging of stage Ⅲ or Ⅳ, and higher levels of IL-6 level, low adiposity index, low skeletal muscle index, and malnutrition (P<0.05), and lower levels of total protein, albumin, hemoglobin level, and prognostic nutritional index (PNI) (P<0.05). There was no significant difference in age, gender, height, weight, BMI and disease type between the two groups (P>0.05). Logistic regression analysis showed that smoking index ≥400, advanced disease stage, IL-6 ≥3.775 ng/L, and PNI <46.43 were independent risk factors for high energy metabolism in lung cancer patients. The AUC of the ROC curve for the established prediction model of high energy metabolism in lung cancer patients was 0.834(95%CI 0.763-0.904). Conclusion The high energy metabolic risk prediction model of lung cancer patients established in this study has good fit and prediction efficiency.

Objective To explore the application value of pathogen-targeted next-generation sequencing (tNGS) technology in patients with suspected pulmonary infections. Methods A retrospective analysis was conducted on the clinical data of 80 patients with suspected pulmonary infections admitted to the Department of Respiratory and Critical Care Medicine at the Third People's Hospital of Hubei Province from January 2021 to July 2023. All patients underwent bronchoalveolar lavage fluid (BALF) tNGS and conventional pathogen detection. Demographic characteristics of the patients were analyzed, and the distribution of pathogens detected by tNGS and conventional methods were compared. The clinical data of patients diagnosed with single pulmonary infections and those with mixed infections were also compared. Results Among the 80 patients, 74 were diagnosed with infections. Most of the infected patients had underlying conditions, mainly chronic heart disease (42.5%), chronic respiratory disease (35%), and diabetes (20%). The tNGS test results led to changes in treatment strategy for 35 patients (43.8%). A total of 45 types of pathogens were detected, with 169 strains identified by tNGS and 63 strains by conventional methods. Within pathogens detected by both methods, bacteria were detected the most. The order of pathogen types detected by tNGS was bacteria > viruses > fungi > atypical pathogens > Mycobacterium tuberculosis. The order of pathogen types detected by conventional methods was fungi > viruses > bacteria > atypical pathogens > Mycobacterium tuberculosis. The consistency between the two pathogen detection methods was poor (kappa value 0.172, P=0.020). The number of positive cases and the positive detection rates for bacteria, viruses, and atypical pathogens detected by tNGS were significantly higher than those of conventional methods (P<0.05), but there was no statistically significant difference in the positive detection rates for fungi and Mycobacterium tuberculosis between the two methods (P>0.05). Using clinical diagnosis as the gold standard, the sensitivity of tNGS detection was significantly higher than that of conventional methods (P=0.026), while there was no statistically significant difference in specificity between the two methods (P>0.05). Among the 74 confirmed pulmonary infection cases, 6 had no clear pathogen, 23 had single infections, and 45 had mixed infections. Among the mixed infections, the most common combination was bacterial-viral mixed infections (12/45, 26.7%). The mortality rate and hospitalization duration of patients with mixed infections were significantly higher than those with single infections (P<0.05); there were no statistically significant differences in gender, age, underlying conditions, white blood cell count, and neutrophil percentage between the two groups (P>0.05). Conclusions tNGS technology has higher pathogen detection sensitivity compared to conventional methods, especially for bacteria, viruses, atypical pathogens, and rare pathogens. This technology is beneficial for identifying mixed infections and can serve as a supplement to conventional pathogen detection methods in clinical practice.

Post-stroke cognitive impairment (PSCI) is a prevalent functional impairments following stroke that seriously affects patients' quality of life and daily activities. Studies indicate a close relationship between intestinal microflora dysbiosis and central nervous system diseases. Intestinal microflora profoundly impacts on human physiological health, contributing to the stability of nervous, metabolic and immune systems through regulation of the gut-brain axis. An increasing number of studies confirmed the important role of the gut microbiome-gut-brain axis in the occurrence and development of stroke and its associated PSCI, and regulation of microbiome-gut-brain could be potential target to treatment of PSCI. This review summarizes research progress on gut microbiome-gut-brain axis and PSCI to provide a reference for exploration of related mechanisms and clinical prevention and treatment strategies.