Objective: To elucidate the effect of targeting acidic nucleoplasmic DNA-binding protein 1 (And-1) on the sensitivity of ovarian cancer cells to the PARP inhibitor niraparib and explore its potential underlying mechanism. Methods: First, And-1 expression were analyzed in ovarian cancer tissues using data from the GEO and TCGA databases, and its impact on patient survival outcomes were examined. Next, ovarian cancer cell lines with stable And-1 knockdown were generated via shRNA technology, and cell proliferation was assessed using growth curves. Cellular sensitivity to niraparib was then measured with CCK-8 assay and further validated through colony formation assays. Additionally, Western blot was performed to assess the expression of the apoptosis-related protein cleaved PARP1 following niraparib treatment, and flow cytometry was used to assess apoptosis rates. Results: And-1 expression was significantly elevated in ovarian cancer tissues compared to normal tissues, and higher And-1 levels were significantly associated with worse prognosis (HR>1,P<0.05). In cell-based experiments, And-1 depletion in ovarian cancer cell lines A2780 and OVCAR3 led to a significant reduction in cell proliferation (P<0.05). Moreover, And-1 depletion markedly lowered the IC50 value of niraparib (P<0.05), and colony formation assays confirmed increased sensitivity to niraparib (P<0.05). Additionally, Western blot analysis showed a marked upregulation of cleaved PARP1 (P<0.05), while flow cytometry revealed a substantial increase in apoptosis (P<0.05). Conclusion: Targeting And-1 enhanced the sensitivity of ovarian cancer cells to niraparib, likely through the promotion of apoptosis.
Objective: To investigate the adverse effect of chemotherapy and immunotherapy on hematopoietic stem/progenitor (Lin-Sca-1+c-kit+, LSK) cells and mature blood cells in the blood system, and further explore whether this effect is exacerbated during chemotherapy combined with immunotherapy. Methods: Based on their distinct surface markers, LSK cells are classified into hematopoietic stem cells (HSCs) and multipotent progenitor cells 1, 2, and 3 (MPP1, MPP2, MPP3). (1) Flow cytometry analysis of the proportion of LSK in the bone marrow of wild-type mice treated with chemotherapy drug carboplatin; (2) Construct a melanoma tumor bearing mouse model, with a control group (injected with PBS) and experimental groups (carboplatin group: injected with carboplatin/trilacilib; PD-1 antibody group: injected with PD-1 antibody; combined treatment group: injected with carboplatin/Trilacilib and PD-1 antibody). Flow cytometry was used to detect the number of LSK in the bone marrow; (3) The automated hematology analyzer and flow cytometry were used to detect the number of red blood cells, white blood cells, and platelets in the peripheral blood of mice, as well as the proportion of B cells, T cells, and myeloid cells. The number of LSK cells in the spleen and the proportion and number of cells at different stages of erythroid differentiation in the bone marrow were also measured. Results: (1)Carboplatin treatment significantly reduced the proportion of LSK cells in mouse bone marrow (P<0.05) and significantly increased the proportion of peripheral blood B cells (P<0.05); (2) PD-1 antibody treatment resulted in significant increase in the proportion of peripheral blood T cells (P<0.05) and a significant decrease in the proportion of peripheral blood myeloid cells (P<0.05). Combined treatment resulted in a significant decrease in the number of peripheral blood white blood cells (P<0.01) and a significant decrease in the proportion of peripheral blood myeloid cells (P<0.05). PD-1 antibody treatment and combined treatment resulted in significant increase in the proportion and number of LSK cells in the spleen (P<0.05); (3) In the carboplatin group, the number of LSK cells in mice was significantly reduced (P<0.05), the number of HSCs was significantly reduced (P<0.05), the number of MPP1 cells was significantly reduced (P<0.05), the number of MPP2 cells was significantly increased (P<0.05), and the number of MPP3 cells was significantly reduced (P<0.05). In the combined treatment group, there was no significant difference in the number of LSK cells in mice (P>0.05), the number of MPP1 cells was significantly reduced (P<0.05), the number of MPP2 cells was significantly increased (P<0.05), and there was no significant difference in the number of MPP3 cells (P>0.05); (4) The combined treatment reduced the number of mature red blood cells (P<0.05) and significant increased the number of upstream red blood cells (P<0.01). Conclusion: Carboplatin treatment inhibits the number of bone marrow LSK cells of wild type mice; For tumor bearing mice, PD-1 antibody treatment increases the proportion of peripheral blood T cells; The combination therapy of carboplatin and PD-1 antibody did not significantly increase the inhibition of carboplatin on the number of LSK cells in the bone marrow. Combined treatment significantly inhibits the number of peripheral blood myeloid cells and white blood cells, promotes extramedullary hematopoiesis, suppresses the number of mature red blood cells in the bone marrow, and promotes significant increase in the number of upstream cells.
Objective: To develop and validate a ferroptosis-related genes (FRGs) model for predicting treatment response in lung adenocarcinoma (LUAD) patients. Methods: Multi-omics data from LUAD and adjacent normal tissues were integrated to identify differentially expressed genes (DEGs) and FRGs. A risk model was constructed utilizing optimized strategy of Cox-LASSO regression analyses. The performance of this model was validated through independent GEO datasets and experimental verification employing quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC). LUAD patients were stratified into high-risk and low-risk cohorts through risk score, with subsequent comparison of inter-group variations in biological processes, immunophenotypic profiles, therapeutic responsiveness and clinical outcomes. In silico analysis was performed to identify potential therapeutic targets for high-risk patients. Results: A five-gene risk model (SLC2A1, TIMP1, CAV1, PECAM1 and COL5A1) was established and validated. High-risk patients exhibited altered expression of these genes (SLC2A1, TIMP1 and COL5A1 upregulated; CAV1 and PECAM1 downregulated), impaired anti-tumor immunity, and reduced response to both chemotherapy and immunotherapy, leading to poorer prognosis. Analysis suggested HDAC1/2 as a potential therapeutic target to overcome treatment resistance in high-risk group. Conclusion: The FRGs-related model (SLC2A1, TIMP1, COL5A1, CAV1 and PECAM1) effectively predicts therapeutic sensitivity in LUAD patients, while unveiling the resistance to standard therapeutic regimens and the clinical translational potential of targeting HDAC1/2 in high-risk LUAD patients.
Objective: To investigate the expression and potential functions of circular RNAs (circRNAs) in hypoxia-induced human retinal microvascular endothelial cells (HRMECs), aiming to provide new insights into the pathogenesis of retinopathy of prematurity (ROP). Methods: HRMECs were isolated and cultured, and a hypoxic cell model of ROP was established. Differential expression profiles of circRNAs were analyzed using whole-transcriptome sequencing. Eight differentially expressed circRNAs (DE-circRNAs) were selected and validated via quantitative reverse transcription polymerase chain reaction (qRT-PCR). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed to explore potential biological functions. A circRNA-miRNA-mRNA regulatory network was constructed using bioinformatics tools. Results: A total of 1,714 circular RNAs (circRNAs) were differentially expressed between normoxic and hypoxia-induced HRMECs, including 899 upregulated and 815 downregulated circRNAs (q<0.05, fold change >2). Gene Ontology (GO) analysis revealed that the differentially expressed circRNAs (DE-circRNAs) were primarily enriched in biological processes such as positive regulation of apoptotic signaling pathways. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis indicated that these DE-circRNAs were mainly involved in tumor-related signaling pathways and the tumor necrosis factor signaling pathway. The circRNA-miRNA-mRNA regulatory network analysis suggested that hsa_circ_0140253 may play a potential regulatory role in the pathogenesis of retinopathy of prematurity (ROP) through the hsa-miR-210-3p/ERFE and hsa-miR-210-3p/PPARGC1A axes. Conclusion: This study reveals the differential expression profile of circRNAs in the hypoxia-induced HRMEC model and suggests that hsa_circ_0140253 may participate in the development of ROP via a competing endogenous RNA (ceRNA) mechanism.
Objective: This study aimed to identify and optimize the active components of Lingguizhugan Decoction (LGZGD) for treating heart failure (HF) using network pharmacology and algorithmic models, and to explore their underlying molecular mechanisms. Methods: Pathogenic genes related to HF were retrieved from the DisGeNet database to construct a weighted gene interaction network. Chemical constituents of LGZGD were obtained from traditional Chinese medicine databases, and active compounds were screened based on ADMET (absorption, distribution, metabolism, excretion, and toxicity) properties and literature evidence. The prediction of compound targets and the construction of the component-target network were both conducted based on the SwissTargetPrediction platform. A node importance evaluation model and cumulative contribution rate algorithm were applied to identify effector space proteins, leading to the selection of a core functional component group. To experimentally validate the findings, an isoproterenol (ISO)-induced myocardial cell injury model was used to evaluate seven core components. Quantitative real-time PCR (qRT-PCR) was performed to measure mRNA expression levels of atrial natriuretic peptide (ANP) and brain natriuretic peptide (BNP), while Western blot was conducted to verify pathway-related proteins. Results: From an initial pool of 662 compounds, 303 active components were identified. Network analysis identified 233 effector space proteins and a core functional component group. These core components exerted therapeutic effects by modulating key signaling pathways, including PI3K/Akt, MAPK, and cAMP. Among them, benzyl acetate, ethyl benzoate, and hydrocinnamic acid significantly suppressed ANP and BNP mRNA levels in injured cardiomyocytes, downregulated PI3K/Akt and MAPK pathways, and activated the cAMP pathway. Conclusion: The optimized network pharmacology model successfully identified core components of LGZGD for HF treatment. These findings suggest that the therapeutic effects of LGZGD were mediated through coordinated modulation of multiple signaling pathways, offering valuable insight into the mechanistic basis of traditional Chinese medicine and supporting the optimization of its formulations.
Objective: Three primary culture methods for dental follicle stem cells' production efficiency and biological characteristics were compared. Methods: Dental follicle stem cells (DFSCs) were isolated using enzymatic digestion (Ed), tissue explant (Te), and hybrid methods(Td). The economic cost, time cost, primary DFSC yield, and biological characteristics were compared among the three groups. Results: Regarding production efficiency, Td demonstrated superior primary cell yield and cost-effectiveness but required longer operational time. Te method was simpler but yielded fewer primary cells. No significant differences were observed in surface markers, migration rates, or multidifferentiation potential among the groups (P>0.05). However, Td group exhibited significantly stronger proliferative capacity compared to Te and Ed (P<0.05), with Ed showing the weakest proliferation (P<0.05). Conclusion: Td is best for large-scale research like cell therapy and tissue engineering due to its high yield. Te is simple and preserves cell viability, making it suitable for small-scale experiments. Ed is used when moderate cell quantities are needed and cell quality standards are not strict. The findings of this study can serve as a reference for optimizing the culture method of DFSC.
Objective: To explore the potential targets and mechanism of procyanidin B2 (PB2) against prostate cancer (Pca) based on network pharmacology. Methods: The core targets of PB2 against Pca were predicted by network pharmacology, and the biological processes and key signaling pathways were enriched and analyzed. CCK-8 method was used to detect the anti-Pca activity of PB2 in vitro. The anti-Pca effect of PB2 was characterized by colony formation and scratch test. The core targets and key signaling pathways predicted by network pharmacology were verified by RT-qPCR and Western blot. Results: A total of 30 intersection targets of PB2 and Pca were screened based on network pharmacology. Among them, janus kinase 2 (JAK2), albumin (ALB), matrix metalloproteinase 9 (MMP9) and phosphoinositide-3-kinase regulatory subunit 1 (PIK3R1) were the key targets of PB2 in the treatment of Pca. MAPK-JAK2/STAT3 is the key pathway. The results of in vitro experiments showed that PB2 could significantly inhibit the proliferation, colony formation and migration of human prostate cancer cells (LNCAP cells). PB2 also significantly down-regulated the mRNA expression levels of core targets ALB, MMP9, PIK3R1 and JAK2 and inhibited the activation of MAPK-JAK2/STAT3 pathway. Conclusion: PB2 may treat prostate cancer through multi-target and multi-pathway synergy; pB2 can effectively inhibit the proliferation and migration of LNCAP cells, down-regulate the mRNA expression of core targets and inhibit the activation of key signaling pathways, thereby exerting an anti-Pca effect.
Objective: Network pharmacology, molecular docking and animal experiments were used to explore the mechanism of action of Wenyang Jiedu Granules on acute lung injury associated with influenza A virus infection. Methods: The active ingredients of Wenyang Jiedu Granules were collected based on the herbal medicine group review (HERB) database, and their ingredient targets were predicted using the SwissTargetPrediction database. Influenza A virus infection disease targets were retrieved using the GeneCards, DisGennet, Pharmgkb, and CTD databases to locate intersection targets. The STRING platform was used to draw the PPI network diagram, and the CytoScape 3.9.1 software was used to construct the TCM-ingredient-target network, and gene ontology (GO) analysis and Kyoto encyclopedia of genes and genomes (KEGG) analysis were performed. The AutoDockTools1.5.7 software was used for molecular docking to explore the potential mechanism, and the influenza A virus-infected mouse model was used for drug effectiveness and mechanism verification. Results: The core active ingredients of Wenyang Jiedu Granules in the treatment of acute lung injury are Tangeretin, Magnococline, Sexangularetin, Retusin, Nomilin, etc, and the core targets are JUN and TP53., SRC, STAT3, EGFR and PIK3CA, etc. GO analysis mainly involved biological processes such as phosphorylation, cytokine-mediated signaling pathways, while KEGG analysis mainly involves signaling pathways such as phosphatidylinositol-3-kinase (PI3K)-AKT signaling pathway. Molecular docking showed that the key components had good binding activity with the core targets. The animal experimental results of this study showed that Wenyang Jiedu Granules can alleviate the pathological damage of mouse lung tissue caused by influenza A virus infection and reduce the expression of inflammatory factors(IL-6, IL-1β, TNF-α, IFN-γ) in lung tissue. Western blot results showed that Wenyang Jiedu Granules could inhibit the expression of PI3K/AKT/NF-κB signaling pathway related proteins. Conclusion: Wenyang Jiedu Granules may inhibit PI3K/AKT/NF-κB signaling pathway through ingredients such as nocturine and nomiline in the treatment of influenza A virus infection-related acute lung injury.
Objective: To establish an individualized model using conventional laboratory indicators combined with tumor markers, leading to the development of a new method for screening and auxiliary diagnosis of lung cancer metastasis. Methods: Clinical data of 218 patients diagnosed with lung cancer at the Affiliated Hospital of Southwest Medical University from 2013 to 2023 were screened and randomly divided into a training set and a validation set. Based on the training set, LASSO regression and multivariate Logistic regression were used to analyze the independent predictors of lung cancer metastasis, and a nomogram model was constructed. The consistency index(C-index), Area Under the Curve (AUC), calibration curve and decision curve analysis (DCA) were used to evaluate the discrimination, calibration, predictive and clinical utility of the model. Results: Multivariate Logistic regression analysis showed that carcinoembryonic antigen (CEA, P<0.001), neuron-specific enolase (NSE, P=0.006), cytokeratin 19 fragment (CYFRA21-1, P=0.025), alanine aminotransferase (ALT, P=0.002), total protein (TP,P=0.006), lymphocyte (LYM, P=0.006), fibrinogen (FIB,P=0.027) were independent predictors of lung cancer metastasis. The C-index of training set and validation set were 0.900 and 0.831, respectively. Calibration and DCA curves showed good predictive performance of the model. Conclusion: CEA, NSE, CYFRA21-1, ALT, TP, LYM and FIB are independent predictors of lung cancer metastasis. The individualized nomogram model for lung cancer metastasis using conventional laboratory indicators combined with tumor markers has good clinical prediction performance and application potential.
Objective: Familial platelet disorder with a predisposition to acute myeloid leukemia (FPD/AML) is a rare autosomal dominant disorder caused by runt-related transcription factor 1 (RUNX1) genetic variant. This study aimed to explore the clinical phenotypic and RUNX1 genotypic characteristics of a pediatric patient with FPD/AML, who is a three-day-old male, providing evidences for the diagnosis and management of this condition. Methods: Clinical information of the patient was collected and analyzed. Peripheral blood DNA was extracted from the patient and his parents. Next-generation sequencing (NGS) was used to explore the genetic causes. Minigene splice variant analysis was employed to study the aberrant transcript arising from novel splice-site variant. The pathogenicity was assessed according to American College of Medical Genetics and Genomics guidelines. Results: The patient was admitted due to thrombocytopenia uncovered for two days. Laboratory analysis revealed significant thrombocytopenia and dysplasia of megakaryocytes in the bone marrow. Genetic testing identified a heterozygous RUNX1 variant c.509-2A>G, which was not previously reported in any official literatures. On minigene analysis, this variant resulted in the formation of an aberrant transcript r.509_515del (p.Gly170Alafs*3). The patient was clearly diagnosed with FPD/AML. Symptomatic and supportive therapeutics stabilized the platelet count, which remained below the lower limit. The long-term outcome needed to be followed-up. Conclusion: This study identified a novel RUNX1 splice-site variant c.509-2A>G and confirmed its pathogenic role in FPD/AML by using Minigene splicing assay. The findings expanded the RUNX1 variant spectrum and had reference value for the diagnosis and management of FPD/AML.
Objective: The present study aimed to analyze the human papillomavirus (HPV) infection rate and subtype infection among 14 794 samples in Guangzhou, and to explore the association between HPV infection and ThinPrep Cytology Test (TCT). Furthermore, it evaluated the epidemiological features of HPV infection in Guangzhou, Guangdong Province, provided epidemiological insights into HPV infection among men, and assessed the significance of combined HPV and TCT testing in cervical cancer screening for women and in devising vaccination strategies against HPV. Methods: Sample data from 14,794 individuals who underwent genotyping for 37 HPV subtypes at the Second Affiliated Hospital of Guangzhou University of Chinese Medicine between January 2023 and December 2023 were collected. Concurrent analysis of TCT screening results was conducted to statistically examine the HPV infection characteristics across different ages, genders and seasons, as well as the relationship between single HPV infection, multiple HPV infection and TCT results. Results: The overall HPV infection rate stood at 29.42%. The most prevalent HPV subtype infections were HPV52 (21.55%), HPV58 (10.00%), HPV16 (9.17%), HPV51 (8.37%), HPV39 (7.56%), HPV61 (7.34%), and HPV53 (7.01%). The infection rate was 29.03% among women and 43.25% among men. The HPV infection rate exhibited a close correlation with age distribution, with statistically significant variations noted in the overall infection rate, single infection rate, and multiple mixed infection rate across different age groups ( X2=144.641, 30.797; P<0.001). Seasonal variations in infection rates were also observed, with rates of 28.03% in spring, 28.58% in summer, 29.56% in autumn, and 32.26% in winter. Among single infections, the highest infection rate of high-risk subtypes occurred in summer (43.53%), while among multiple infections, the higher infection rate was noted in autumn (35.02%). The TCT groups were predominantly characterized by single infections. Significant differences were discernible between single infections and multiple infections within each TCT group ( X2=90.497, P<0.001). Notably, TCT results also varied significantly among different age groups ( X2=32.871, P<0.001). Conclusion: The present study illuminated the epidemiological profile of HPV infection in the Guangzhou area, with an overall infection rate of 29.42%. HPV52, HPV58, and HPV16 emerged as the most frequent types. The overall HPV infection rate among men (43.25%) surpassed that among women (29.03%). The HPV infection rate showed a strong correlation with age distribution and was higher in summer and autumn compared to spring and winter. Additionally, this study identified significant differences in HPV infection rates among TCT groups and TCT results across different age groups, furnishing a scientific rationale for early HPV screening and presenting novel perspectives on cervical cancer prevention and treatment. In summary, this study contributes vital epidemiology data on HPV infection in Guangzhou, providing a scientific foundation for the formulation of targeted HPV prevention and control strategies and enhancement of cervical cancer diagnosis and treatment. It underscores the crucial role of male HPV infection in HPV prevention and control efforts.
Objective: To explore the intervention effects of open-kinetic chain and closed-kinetic chain exercises on patients with patellofemoral pain (PFP). Methods: A total of 46 patients with patellofemoral pain were randomly divided into the open-kinetic chain group and the closed-kinetic chain group, with 23 cases in each group. The open-kinetic chain group received open-kinetic chain action movement of the hip and knee joint muscle groups, while the closed-kinetic chain group received corresponding closed-kinetic chain action movement. Before and after the training, both groups of patients underwent visual analog scale (VAS) for pain, kujala patellofemoral scale (KPS) for knee joint function, and Tampa scale of kinesiophobia (TSK) assessments, and the hip and knee joint angles and muscle activation ratios during the step-down test were measured. Results: After 6 weeks of training, both groups showed significant improvements in VAS, TSK, and KPS scores (P<0.05). The VAS and TSK scores of the closed-kinetic chain group were lower than those of the open-kinetic chain group (P<0.05), and the KPS score was significantly higher than that of the open-kinetic chain group (P<0.05). The knee flexion angle, vastus lateralis activation degree, and the ratio of vastus medialis to vastus lateralis activation time in the open-kinetic chain group increased (P<0.05), while the ratio of vastus medialis to vastus lateralis activation degree decreased (P<0.05). In the closed-kinetic chain group, the hip flexion angle and gluteus maximus activation degree increased (P<0.05), and the knee valgus angle decreased (P<0.05).KPS scores. The open-chain group experienced significant increases in knee flexion angle, activation of the vastus lateralis, and the activation time ratio of the vastus medialis/vastus lateralis, while the activation ratio of the vastus medialis/vastus lateralis significantly decreased. The closed-chain group showed significant improvements in hip flexion angle, knee varus angle, and gluteus maximus activation. Conclusion: Both open-kinetic chain and closed-kinetic chain exercises are beneficial for alleviating pain, reducing the degree of movement fear, and improving knee joint function in PFP patients. However, from the perspectives of self-assessment scales and biomechanics, closed-kinetic chain exercises have better short-term intervention effects on PFP patients.
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