Combinatorial Chemistry & High Throughput Screening - Online First

Pulsatillae radix (PR), a medicinal root plant and a well-known Chinese herbal remedy, is primarily used for its heat-clearing, detoxifying, blood-cooling, and antiinflammatory properties.

The constituents of PR were systematically analyzed using UHPLC-Q-TOF-MS/MS. Potential targets of active components were identified via the SwissTargetPrediction and PharmMapper databases, while (ulcerative colitis) UC-related disease targets were retrieved from GeneCard and other relevant databases. Overlapping targets between PR and UC were determined using Venn analysis. The STRING database was employed to generate a PPI network for the intersecting targets, and core targets were identified using the CytoNCA plugin. GO and KEGG pathway enrichment analyses were conducted using the DAVID platform. Lastly, molecular docking of key components with target proteins was carried out using PyMOL.

A total of 27 active compounds, 237 drug targets, and 4622 disease targets were identified. Intersection analysis revealed 141 shared targets, while the PPI network identified 10 hub targets. GO and KEGG enrichment analyses indicated that the hub targets were primarily associated with phosphorylation, cytoplasmic functions, nuclear receptor activity, as well as pathways related to the AGE-RAGE products signaling, TCR signaling, lipid and cholesterol metabolism, and various cancer-related pathways. Molecular docking experiments demonstrated that (+)- pinoresinol, cichoric acid, β-ecdysone, pulsatilla saponin D, 23-HBA, and AB4 exhibited stable binding to PIK3R1, TLR4, and ESR1, with AB4 forming the most stable complex with ESR1.

PR exerts therapeutic effects on UC through the synergistic actions of multi-components (AB4, 23-HBA), multi-targets (ESR1, TLR4, PIK3R1), and multi-pathways (AGE-RAGE, TCR).

Observational studies have shown a link between constipation (CN) and psychiatric disorders, including Schizophrenia (SP), Bipolar disorder (BD), Schizoaffective disorder (SD), and Parkinson’s disease (PD). However, it is still unknown whether CN affects the occurrence and development of psychiatric disorders or whether psychiatric disorders cause the occurrence and development of CN. Therefore, this study used Mendelian randomization (MR) analysis to evaluate the relationship between CN and psychiatric disorders.

We used genome-wide association studies (GWAS) to assess the relationship between constipation (N = 411, 623) and four psychiatric disorders, including SP (N = 77, 096), BD (N = 51, 710), SD (N = 210, 962), PD (N = 482, 730), using bidirectional MR analysis. Inverse variance weighting (IVW), MR Egger (ME) and Weighted median (WM) were used as causal analysis methods. Cochran's Q test, funnel plot, MR Egger intercept test and Leave‐one‐out analysis were used to detect sensitivity. Confounding factors were analyzed and eliminated by LDtrait to avoid influencing the final MR Analysis result.

The results of positive MR analysis indicated that there was no evidence of influence of constipation on SP (OR 1.043, 95%CI 0.946 - 1.149, P value = 0.398), BD (OR 1.114, 95%CI 0.995 - 1.248, P value = 0.062), SD (OR 0.934, 95%CI 0.674 - 1.294, P value = 0.682) and PD (OR 1.118, 95%CI 0.918 - 1.361, P value = 0.269) under gene prediction. Reverse MR analysis suggested that SP (OR 1.030, 95% CI 1.001-1.060, P value = 0.042) had a causal relationship with constipation. BD (OR 0.993, 95% CI 0.962-1.025, P value = 0.664), SD (OR 1.021, 95% CI 0.984-1.059, P value = 0.265) and PD (OR 1.004, 95% CI 0.974-1.035, P value = 0.790) were not associated with CN.

There was a positive association between SP and CN. CN may have no exact causal relationship with BD, SD and PD, and the interaction mechanism between these diseases needs to be further explored.

Bladder cancer (BC) is one of the most common urological malignancies, ranking as the eleventh most common cause of cancer-related deaths worldwide. The lack of specific and sensitive prognostic biomarkers presents a significant challenge in the early diagnosis and treatment of BC.

This study utilizes the Gene Expression Omnibus (GEO) dataset GSE13507 and the Cancer Genome Atlas (TCGA) database to screen differentially expressed genes related to BC. By using Weighted Gene Co-expression Network Analysis (WGCNA), two modules associated with BC were investigated in GSE13507 and TCGA. Hub genes were identified through Protein-Protein Interaction (PPI) network analysis, and their functions were validated through multiple approaches, including Gene Expression Profiling Interactive Analysis (GEPIA), Western Blotting (WB) assay, Human Protein Atlas (HPA), Oncomine analysis, and quantitative Real-Time PCR (qRT-PCR) analysis. Additionally, miRNAs associated with hub gene expression were identified using various databases to predict the progression and prognosis of BC.

WGCNA and a differential gene expression analysis identified 171 common genes as target genes. Ten genes (MYH11, ACTA2, TPM2, ACTG2, CALD1, MYL9, TPM1, MYLK, SORBS1, and LMOD1) were identified using the PPI tool. The CALD1 and MYLK genes showed a significant prognostic value for overall survival and disease-free survival in patients with BC. CALD1 and MYLK expression levels were significantly lower in BC tissues than in normal tissues. Furthermore, miR-155 showed a significant positive correlation with MYLK.

This study established MYLK as a direct target gene of miR-155, functioning as an actionable survival-related gene correlated with BC development.

This study aims to clarify the role of kelch like family member 17 (KLHL17) in cervical cancer (CESC) is unclear.

To clarify this uncertainty, our research employed bioinformatics analysis coupled with experimental corroboration.

We utilized the Cancer Genome Atlas (TCGA) database to assess the expression of KLHL17 in various cancers, specifically CESC, and to explore its association with clinical characteristics, diagnostic utility, and prognostic significance in CESC. The current investigation delved into the potential regulatory pathways related to KLHL17, examining its connection with the infiltration of immune cells, the expression of immune checkpoint genes, the status of microsatellite instability (MSI), and the efficacy of diverse therapeutic agents in CESC. The research analyzed KLHL17 expression patterns using single-cell sequencing data from CESC samples and investigated the genetic variations of KLHL17 within this context. KLHL17 expression was validated using GSE145372. The presence and levels of KLHL17 in different cell lines were validated through quantitative real-time PCR (qRT-PCR) assays.

KLHL17 exhibited irregular expression profiles across various cancer types, including CESC. Furthermore, increased KLHL17 levels in CESC patients were significantly associated with a lower progression-free survival (PFS) rate (hazard ratio: 1.62; 95% confidence interval: 1.01–2.60, p = 0.044). Moreover, KLHL17 expression emerged as a distinct prognostic indicator for CESC patients (p = 0.031). It has been associated with various biological pathways, such as cytokine-cytokine receptor interaction, primary immunodeficiency, cell adhesion molecules (CAMs), chemokine signaling pathway, steroid hormone biosynthesis, and others. The expression levels of KLHL17 were found to correlate with the presence of immune cells, the expression of immune checkpoint genes, and the status of MSI within CESC. Furthermore, KLHL17 expression exhibited a significant and inverse correlation with XMD15-27, rTRAIL, Paclitaxel, tp4ek, and tp4ek-k6. Furthermore, KLHL17 was found to be significantly positively regulated in CESC cell lines.

The findings suggest that KLHL17 is involved in the progression of CESC and may serve as a potential prognostic marker and therapeutic target. KLHL17's association with immune cell infiltration and immune checkpoint genes indicates a role in immuneevasion. Future research should focus on validating these findings through independent datasets and experimental studies to elucidate the molecular mechanisms underlying KLHL17's role in CESC progression and immune regulation.

KLHL17 is a promising prognostic marker and potential therapeutic target in CESC.

For clarifying the “multi genes and multi targets” characteristic of the treatment of Erxia Decoction (EXD), the aim of this study was to employ network pharmacology technology to perform cluster analysis on selected EXD targets.

EXD, a famous Chinese herbal prescription, consisting of Pinelliae Rhizoma (PR) and Prunellae Spica (PS), was mainly used to treat sleep disorder (SLD).

Using network pharmacology combined with proteomics to find out the main active components and core targets of EXD in the treatment of SLD.

By constructing the network of drug–component–target, the key protein targets of EXD for the treatment of SLD were screened. Then the interaction of the main active components of EXD and predicted candidate targets were verified. Then the proteomic analysis was used to screen the core targets in BV2 cells treated with EXD or the chemical ingredients, and the expression level was validated by Western blotting. Finally, molecular docking was used to further evaluate the mechanism of the action of the main ingredients and the core targets.

The 24 components of EXD mainly participate in the SLD treatment process by acting on 15 important key genes, and the core signal pathways were identified in the process of the action of EXD in treating SLD. Four key ingredients and five core targets were revealed from the results of network pharmacological analysis combination with proteomics, and then the AKT1 protein as a key target was validated by PCR and Western blotting.

This study preliminarily revealed EXD, morin (MOR) and quercetin (QUE) mainly inhibited the AKT1 core targets for the treatment of SLD using the network pharmacological analysis, proteomics, Western blotting and molecular docking.

The results elucidated partly the molecular mechanism and provided clues and a basis for further research.

Immunoglobulin A (IgA) Nephropathy (IgAN) is characterized by pIgA1 dep-osition in the glomerular mesangium, resulting in podocyte Epithelial-to-Mesenchymal Transi-tion (EMT). Traditional Chinese Medicines (TCMs) have been used for the treatment of IgAN for several years and have demonstrated positive efficacy. The present study aimed to establish a High-Content Screening (HCS) method for identifying wind-dampness dispelling TCM extracts that can mitigate podocyte EMT induced by pIgA1.

IgA1 from IgAN patients was used to establish a podocyte EMT model. The expression of EMT markers, including desmin, podocalyxin, and podocin, was assessed by Immunocytofluorescence (IF). An image-based HCS method was established to identify wind- dampness dispelling TCMs with the anti-EMT activity of podocyte EMT by automatic acquisi- tion and processing of dual-fluorescent labeled images.

A total of 21 wind-dampness-dispelling TCM extracts were screened using the HCS system, leading to the identification of eight wind-dampness-dispelling TCM extracts exhibiting anti-EMT activity. These eight extracts inhibited the pIgA-induced expression of desmin while upregulating the expression of podocalyxin and podocin.

By quantifying the changes in the expression of EMT markers during pIgA1- induced EMT, this study successfully identified wind-dampness dispelling TCM extracts with anti-EMT properties using an HCS system. In addition, the proposed approach presents a novel avenue for the identification of wind-dampness-dispelling TCMs for the treatment of pIgA1- induced EMT.

Allergic asthma is an inflammatory disease of the airways that causes great distress to the patient's normal life. Astragalus Polysaccharide (APS) is the main active ingredient in the traditional Chinese medicine Astragalus mongholicus Bunge, which has the effect of regulating immune function.

This study aimed to evaluate the effect of APS on allergic asthma and investigate its potential mechanism of action.

This study utilized network pharmacology to predict the relevant targets and signaling pathways of APS treatment for allergic asthma. Subsequently, an animal model was established using Ovalbumin (OVA) induction. The efficacy of APS was verified using histopathologic staining and Airway Hyperresponsiveness (AHR) assay. Signaling pathways were examined using Western Blot (WB). Finally, bioinformatics analysis was utilized to explore the correlation between the progression of allergic asthma and signaling pathways.

Network pharmacology analysis identified 15 intersection targets significantly enriched in the PI3K/AKT signaling pathway. The results of molecular docking showed that small molecule drugs have a strong binding ability to target proteins. The experiments confirmed APS successfully suppressed the pathological symptoms in allergic asthma model mice. Subsequently, WB provided evidence supporting that APS has potential therapeutic effects mediated through the PI3K/AKT signaling pathway. The bioinformatics results confirmed that disease progression in allergic asthma patients does correlate with the PI3K/AKT signaling pathway.

Our study suggests that APS may treat allergic asthma by targeting the PI3K/AKT signaling pathway. This provides a basis for preliminary research on the clinical application of APS for treating allergic asthma.

Our objective is to assess the therapeutic impact of HEC on OAB rats and investigate potential mechanisms.

Overactive bladder (OAB) is a syndrome of urinary storage symptoms characterized by “urinary urgency with or without urinary acute incontinence, usually accompanied by increased daytime and nocturnal urination”, which impacts patients’ quality of life. We found the potential therapeutic impact of HuangE capsules (HEC) on OAB patients through clinical practice. However, the exact effect and mechanism of action remain unclear.

We developed a “drugs- active ingredients- targets- diseases” network and employed the pathway enrichment analysis to identify the potential mechanisms of HEC on OAB. Bladder outlet obstruction (BOO) models and sham-operated ones were established in healthy male Wistar rats through surgical procedures. Following 28 days of continuous gavage administration of HEC, saturated copper sulfate test paper was utilized to quantify the frequency of urination over a 24-hour period. Subsequently, cystostomy was conducted to perform cystometry, and Masson staining was applied to a portion of the bladder tissue. Finally, we investigated the Rho/Rho-kinase pathway's expression and assessed the oxidative stress and inflammatory factor levels in the rat bladder through western blotting and ELISA techniques.

Through network pharmacological analysis, we identified RhoA/Rho-kinase pathway and cytokine including TNF-α, IL-6, SOD and MDA as potential mechanisms of HEC on OAB. The rats in the 2× HuangE group exhibited significantly enhanced urodynamic outcomes and decreased 24-hour urination frequency compared to the model group. Masson staining indicated a decrease in the proportion of collagenous tissue and an improvement in histomorphology. We observed a decrease expression of RhoA, ROCK1, and ROCK2 protein in the bladder tissue of 2× HuangE group rats, along with elevated SOD levels and decreased levels of TNF-α, IL-6, and MDA.

HEC could improve bladder function and morphology in BOO-induced OAB rats by reducing the expression of RhoA, ROCK1, and ROCK2 and lowering levels of oxidative stress and inflammation.

Shenhuang Liuwei powder (SHLWP) is frequently used to treat diabetic ulcers (DUs), but its mechanism of action remains poorly understood. This study aimed to identify the active compounds and mechanisms by which SHLWP alleviates DUs.

The chemical components of SHLWP were analyzed using high-resolution mass spectrometry (HRMS). Network pharmacology based on HRMS data identified SHLWP-associated targets and signaling pathways. Its antibacterial activity was assessed using Kirby-Bauer disc diffusion and minimum inhibitory concentration (MIC) tests. Its in vivo pharmacological effects were evaluated in a streptozotocin-induced diabetic ulcer model in Sprague-Dawley (SD) rats.

Seventy-three components were identified in SHLWP, with key constituents including caffeic acid (13.11 ± 0.14 μg/g), ferulic acid (20.40 ± 0.24 μg/g), quercetin (8.49 ± 0.18 μg/g), luteolin (36.63 ± 0.19 μg/g), apigenin (82.14 ± 1.60 μg/g), and linoleic acid (507.59 ± 1.46 μg/g). SHLWP exhibited strong antibacterial activity against Staphylococcus aureus (MIC = 7.8125 μg/mL), Streptococcus pyogenes (MIC < 3.90625 μg/mL), and Streptococcus epidermidis (MIC < 3.90625 μg/mL). Network pharmacology revealed significant enrichment of the AGE/RAGE, HIF-1, and PI3K-Akt pathways, which was validated in vivo using qPCR, immunohistochemistry, and Western blot.

SHLWP alleviated streptozotocin-induced diabetic ulcers by inhibiting the AGE/RAGE pathway and promoting antibacterial activity and angiogenesis via the PI3K/Akt/eNOS/HIF-1α pathway, providing a biological basis for its therapeutic effects.

The specific role of necroptosis in the pathogenesis of cataracts remains unclear. This study aimed to identify and validate the genes related to necroptosis in the development of cataracts through bioinformatics analysis.

We utilized RNA sequencing data (GSE161701) from the Gene Expression Omnibus (GEO) database and employed R software to perform differential expression analysis of necroptosis-related genes (NRGs) in lens epithelial cells (LECs) under oxidative stress. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to evaluate the functions of necroptosis-related differentially expressed genes (NRDEGs) and their associated pathways. Additionally, a diagnostic model was established using LASSO regression to select hub genes, and protein-protein interaction (PPI) networks, mRNA-miRNA, and mRNA-drug regulatory networks were constructed. Immune infiltration analysis was performed using the xCell and CIBERSORT algorithms, and the differential expression of hub genes was validated in a UVB-induced rat cataract model using RT-qPCR and immunohistochemistry.

The results indicated that oxidative stress promoted necroptosis in LECs, involving 86 NRDEGs and nine hub genes. GO and KEGG analyses revealed significant enrichment in necroptosis-associated pathways. Furthermore, we identified 58 mRNA-miRNA interactions and 131 potential molecular compounds or drugs. The immune infiltration analysis showed that certain immune cells exhibited significantly elevated expression in the cataract group, with notable correlations between some immune cells and hub genes. RT-qPCR and immunohistochemistry confirmed the expression of 9 hub genes and 3 key necroptosis genes. BAX, CXCL1, EPAS1, JUN, LRP1, RBM14, SERTAD1, and TNFAIP3 were highlighted as potential diagnostic and therapeutic targets.

This study identified key NRDEGs involved in the pathogenesis of cataracts under oxidative stress through bioinformatics analyses, potentially providing new targets and research directions for future cataract prevention and treatment.

Chenopodium album Linn. is a nutritionally and pharmacologically significant herb that generally grows in the winter season along with other crops. It is rich in fibers, protein, minerals (Mg, Ca, Fe, K, P, and others), vitamins (ascorbic acids, thiamine, riboflavin, and others), and several other biologically active chemical components like flavonoids, saponins, steroids and many more. In this article, the authors briefly describe and assess the chemistry and pharmacology of this nutritionally significant plant.

This study is based on several literature searches conducted via Google Scholar, Research Gate, PubMed, and many other online sources.

Due to its richness with bioactive phytochemicals, it has become a valuable functional food. C. album has several medicinal properties like antioxidant, antimicrobial, anti-arthritic, anti-diabetic, anti-infection, anti-ulcer, and many others. Even after its rich nutritional values, chemical compositions, and a broad spectrum of pharmacological properties, this is a highly ignored herb worldwide. Therefore, extensive research and awareness regarding the functional role of this herb is needed.

This study aimed to investigate the modulation of intestinal flora by electroacupuncture in a murine ulcerative colitis (UC) model, with a focus on analyzing microbial taxa and identifying key regulatory targets and pathways.

A UC model was established in mice using 5% dextran sodium sulfate (DSS). Electroacupuncture was applied at bilateral “Shangjuxu” (electrostimulation) and “Tianshu” (manual acupuncture) points from days 5–9, while the mesalazine group received 0.5 g/kg/day via gavage. Disease activity index (DAI), colon length, and histopathology (hematoxylin-eosin staining) were evaluated. Intestinal flora composition was analyzed via 16S rDNA sequencing.

Electroacupuncture significantly reduced DAI scores on days 7 and 9 (P < 0.05; P < 0.01) compared to the model group, improved colon morphology, and reduced inflammation. Linear discriminant analysis and Wilcoxon tests revealed an increased abundance of Roseburia and elevated alpha diversity in the electroacupuncture group. Functional prediction demonstrated suppressed RNA transport and glycerophospholipid metabolism in the model group (P < 0.05), which were significantly enhanced post-electroacupuncture (P < 0.01).

Electroacupuncture restored beneficial taxa (e.g., Roseburia) and microbial diversity, suggesting gut homeostasis modulation. Enhanced lipid metabolism and RNA transport pathways may underlie its anti-inflammatory and mucosal repair effects.

Electroacupuncture alleviates UC by modulating the structure and function of intestinal flora, with Roseburia and associated metabolic pathways identified as key targets. These findings highlight the potential of electroacupuncture as a UC therapy.

Atherosclerosis (AS) is a leading cause of cardiovascular diseases, characterized by lipid accumulation in arterial walls. The gene Ephx2, which encodes soluble epoxide hydrolase (sEH), is implicated in AS development, but its precise mechanisms and therapeutic potential are not fully understood.

This study aimed to analyze gene expression data from low-density lipoprotein receptor knockout (LDLR^−/−) and LDLR^−/−sEH^−/− mice to identify significant genes associated with AS.

A directed compound-protein interaction network was constructed based on these genes and related pathways from the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. In the end, through resistance distance (RD) between any two nodes in this network, the Independent Cascade (IC) model was applied to explore Ephx2 mechanisms in AS, such as important Adverse Outcome Pathways (AOPs).

Several AOPs were identified as critical in AS treatment via Ephx2. The key AOPs included inflammatory response and cytokine release, cholesterol deposition and oxidation, disruption of plaque stability, smooth muscle cell proliferation and migration, and platelet activation and coagulation. Within the top AOPs of inflammatory response and cytokine release, potential target genes were identified, such as Mapk3, Pik3cd, Gnai2, Mapk10, Arnt, and RhoA. Critical paths from Ephx2 to these target genes were established, suggesting mechanisms by which Ephx2 may influence AS pathogenesis.

By defining the AS network and corresponding RD, this study elucidates potential mechanisms by which Ephx2 affects AS through specific KEGG pathways, AOPs, and target genes. These findings enhanced the understanding of AS pathogenesis and highlighte potential targets like Mapk3 for developing therapeutic strategies in AS prevention and treatment.

Astragaloside (AST), a natural saponin extracted from Astragalus membranaceus (Fisch.) Bunge., has been consistently utilized in the treatment of rheumatoid arthritis (RA). N6-methyladenosine (m6A), the most prevalent modification of mRNA, is associated with the progression of various diseases, including RA. Nonetheless, the effects of AST on m6A modification in RA remain to be elucidated.

The MH7A cell model was established through induction with TNF-α. The effects of AST on the expression levels of WTAP, BAX, BCL2, and TRAIL-DR4 were evaluated utilizing immunofluorescence, RT-qPCR, and Western blot analysis. Furthermore, CCK-8 and flow cytometry were used to assess MH7A cell viability, cell cycle, apoptosis, and proliferation. Then, the m6A modification of TRAIL-DR4 was elucidated via MeRIP-qPCR.

The optimal dose administration time was 50 μg/mL at 48 h. AST not only reduced the expression levels of WTAP, BCL2, BAX, TRAIL-DR4, and the m6A modification level of TRAIL-DR4 but also significantly enhanced apoptosis in MH7A cell, while inhibiting cell viability and proliferation. Furthermore, AST was capable of reversing the effect on MH7A cell proliferation and apoptosis induced by WTAP overexpression.

This study elucidates the protective role of AST on MH7A cells by attenuating m6A/WTAP-mediated apoptosis, offering novel insights into the mechanisms of AST.

Transcription factors (TF) are the central regulatory hubs of signaling pathways in eukaryotic cells. Here, we explored the abnormal expression of TF in septic-induced lung injury by sequencing.

The levels of target proteins were detected using Western Blot and Elisa. Cell function was evaluated using CCK8 and transwell assays. A double luciferase reporter assay was performed to detect interactions between target molecules.

We found that TF glioma-associated oncogene (GLI) family zinc finger 3 (GLI3) was abnormally low expressed in a lipopolysaccharide (LPS) induced acute lung injury (ALI) cell model. In an in vitro model, GLI3 overexpression promoted the proliferation and migration and inhibited apoptosis of lung epithelial cells in LPS-induced inflammatory environment. Importantly, GLI3 overexpression inhibited the secretion of inflammatory factors IL-1β, IL-6, and TNF-α. Additionally, miR-143-3p inhibited the expression of GLI3. MiR-143-3p inhibitor alleviated the cell damage caused by LPS, while knocking down GLI3 counteracted this effect, indicating that miR-143-3p downregulated GLI3 and inhibited its anti-inflammatory effect. Secreted frizzled related protein-1 (SFRP1) was upregulated in LPS-treated cells and SFRP1 promoter interacted with GLI3, suggesting that SFRP1 was a target of TF GLI3. Co-transfection with GLI3 knockdown and SFRP1 overexpression plasmids attenuated the secretion of inflammatory factors IL-1β, IL-6, and TNF-α caused by GLI3 knockdown in LPS-treated cells, indicating that SFRP1 plays an anti-inflammatory role as a GLI3 target in the ALI cell model.

miR-143-3p caused degradation of GLI3 mRNA and thus inhibited the transcription of SFRP1, leading to decreased proliferation and increased levels of inflammatory factors, providing new potential targets for the clinical diagnosis and treatment of ALI.

Benzoimidazo[1,2-a]pyrimidines are important compounds that have many useful effects in the body. They can help fight cancer, fungal infections, inflammation, and viruses. They can also help with various other health conditions. They can act as antineoplastic, antitubercular, parasitical activity, benzodiazepine receptor agonists, calcium channel blockers, potent P38 MAP kinase inhibitors, TIE-2 and/or VEGFR2 inhibitory activities, protein kinase inhibitors, and T cell activation. There are different methods to make the benzoimidazo[1,2-a]pyrimidines. Some of them dealth with the one-pot threecomponent condensation reactions of β-dicarbonyl compounds, aldehyde and 1H-benzo[d]imidazol-2-amine in the presence of catalyst. Although the synthesis of this group of compounds has been done before, and the products have been identified from the spectroscopic point of view, the kinetics and reaction mechanism have not been investigated. The strength of these calculations is that evaluation of the activation energy of various steps suggests possible mechanisms, probable mechanisms, and valuable synthetic intermediates.

In this report, seven possible mechanisms for synthesizing the benzoimidazo[1,2-a]pyrimidines have been investigated using density functional theory (DFT) at the B3LYP/6-311G** level of theory. Each synthetic route involves condensation of the benzaldehyde, indanedione and 1H-benzo[d]imidazol-2-amine molecules to yield the proposed product. The calculations showed that the suggested method has six steps; its initiation step includes the Knoevenagel reaction between indanedione and aldehyde, and the rate determining state is dehydration in the fifth step.

Six potential pathways for the reaction will occur. Then, we focused on the best pathway and studied it in detail. The ways that three chemicals-indanedione (R1), benzaldehyde (R2), and 1H-benzo[d]imidazol-2-amine (R3) react with each other were studied using ab-initio program by ChemBio3D, Gauss View, and Gaussian 09. The Density Functional Theory (DFT) using the B3LYP basis set was used to improve the arrangement of molecules involved in the three-part creation of a specific compound called 12-phenyl-5H-benzo[4,5]imidazo[1,2-a]indeno[1,2-d]pyrimidin-13(12H)-one (P).

During the study of the six mechanisms, the proposed pathway 2 is the best mechanism for this reaction because its rate-determining step has the lowest activation energy value. This route consists of 6 steps, the fifth step of which is related to the conversion of IM4 to IM5 (relative ∆E: 109.80 Kj/mol), during which a dehydration reaction is performed, and this step occurs by passing through transition state TS5 (Total Energy (Hart./particles: -1194.747403).

Esophageal Squamous Cell Carcinoma (ESCC) remains a significant global health challenge, underscoring the urgent need for the development of innovative therapeutic approaches. Ranunculus ternatus Thunb., a traditional herb, exhibits potential anticancer properties, but its mechanisms against ESCC remain poorly understood. This study integrates network pharmacology and experimental validation to explore the therapeutic effects of the ethyl acetate extract of Ranunculus ternatus Thunb. (RTE).

Potential targets of RTE and ESCC were screened using public databases. A Protein-Protein Interaction (PPI) network was constructed to identify key targets, followed by GO and KEGG pathway enrichment analyses. The predicted mechanisms were validated using in vitro assays, including cell proliferation analysis and western blot assay in ESCC cell lines.

Network pharmacology analysis identified 274 potential targets, with 14 key genes implicated in the therapeutic effects of RTE. GO analysis revealed significant involvement in the inflammatory response and apoptotic signaling pathways. KEGG pathway analysis highlighted the MAPK, Relaxin, and PI3K/Akt signaling pathways as critical mechanisms. In vitro experiments demonstrated that RTE significantly inhibited the proliferation of EC-109 and TE-13 cells by modulating the MAPK/ERK and PI3K/Akt pathways.

The study reveals that active compounds of RTE target MAPK/ERK and PI3K/Akt pathways, aligning with prior evidence. However, future studies should explore animal models to confirm efficacy.

This study provides a comprehensive understanding of the molecular mechanisms underlying the anticancer effects of RTE against ESCC. These findings underscore the potential of RTE as a promising natural compound for ESCC treatment.

The objective of this research is to demonstrate that alignment-free bioinformatics approaches are effective tools for analyzing the similarity and dissimilarity of protein sequences. All numerical parameters representing sequences are expressed analytically, ensuring precision, clarity, and efficient processing, even for large datasets and long sequences. Additionally, a novel approach for identifying previously unknown virus strains is introduced.

A novel approach is proposed, integrating the unique features of our newly developed method, the 20D-Dynamic Representation of Protein Sequences, with the K-means clustering algorithm. The sequences are represented as clouds of material points in a 20-dimensional space (20D-dynamic graphs), with their spatial distribution being unique to each protein sequence. The numerical parameters, referred to as descriptors in molecular similarity theory, represent quantities characteristic of dynamic systems and serve as input data for the K-means clustering algorithm.

Examples of the application of the approach are presented, including projections of the 20D-dynamic graphs onto 3D spaces, which serve as a visual tool for comparing sequences. Additionally, cluster plots for the analyzed sequences are provided using the proposed method.

Combining the 20D-Dynamic Representation of Protein Sequences with an unsupervised machine learning algorithm (K-means clustering) enhances its scalability. This approach is applicable to large datasets without restrictions on sequence length.

It has been demonstrated that the 20D-Dynamic Representation of Protein Sequences, combined with the K-means clustering algorithm, successfully classifies subtypes of influenza A virus strains.

Osteosarcoma (OS), a malignant tumor originating in bone or cartilage, primarily affects children and adolescents. Notably, substantial alterations in mitochondrial energy metabolism have been observed in OS; however, the specific contribution of mitochondrial-related genes (MRGs) to OS pathogenesis and prognosis remains unclear. Herein, we identified novel diagnostic biomarkers associated with mitochondrial-related processes in OS via comprehensive bioinformatics analysis.

OS mRNA expression profiles were retrieved from GSE16088 and GSE19276 databases. Mitochondrial-related differentially expressed genes (MitoDEGs) were identified by integrating differentially expressed analysis with mitochondrial-localized genes. A protein-protein interaction network was constructed, and machine learning algorithms (LASSO regression analysis and SVM-RFE) identified characteristic MitoDEGs. Subsequently, immune cell infiltration, microenvironment analysis, and single-cell RNA sequencing (scRNA-seq) analyzed differences in characteristic MitoDEGs, and RT-PCR was used for in vitro verification of characteristic MitoDEGs.

MitoDEGs in OS were significantly enriched in the pathways associated with mitochondrial function and immune regulation. Two MitoDEGs, UCP2 and PRDX4, were identified via LASSO and SVM-RFE. Correlation analysis demonstrated a close association between UCP2 and PRDX4 expression levels and immune cell infiltration, particularly in CD8+ T and native CD4+ T cells, as observed in both immune cell and scRNA-seq analyses. Furthermore, RT-PCR confirmed the expression levels of UCP and PRDX4 at the cellular level, which was consistent with the bioinformatics results.

This study identified UCP2 and PRDX4 as characteristic MitoDEGs and potential diagnostic biomarkers for OS using machine learning algorithms. These findings provide novel insights into the clinical applications of these biomarkers for OS diagnosis.

This study aimed to assess the safety and effectiveness of carbamazepine in treating trigeminal neuralgia in contrast to gabapentin. Hence, a systematic review and meta-analysis of randomised controlled trials had been carried out.

The relevant studies were searched in PubMed and filtered according to the inclusion and exclusion criteria. Independently, two reviewers chose the studies, evaluated the quality of the investigations, and retrieved the data. RevMan was used for analysis when the data were collected and entered into the data extraction sheet. In addition to heterogeneity, the overall estimate measures were computed as mean differences with a 95% confidence interval for continuous data and relative risk for dichotomous data. To investigate the impact of outliers on the result, a sensitivity analysis was performed. A funnel plot was used to qualitatively evaluate the publishing bias. A total of 1,650 participants from 19 randomised controlled trials were evaluated.

The meta-analysis revealed that the group receiving gabapentin therapy had a similar overall effective rate to the group receiving carbamazepine therapy (OR = 1.94, 95% CI 1.46, 2.57, P = 0.32). Additionally, our meta-analysis revealed that the group receiving gabapentin therapy witnessed a significantly lower risk of adverse reactions than the group receiving carbamazepine therapy (OR= 0.29, 95% CI 0.22, 0.387, P<0.00001).

In summary, the current trials comparing carbamazepine and gabapentin have had inadequate methodological quality. It is not possible to conclude that gabapentin is more effective than carbamazepine in terms of adverse effects based on the evidence that is currently available.

This study aimed to examine the impact of quercetin on a mouse model of endometriosis and elucidate its underlying mechanisms.

An endometriosis model was established using C57BL/6 mice, which were divided into three groups: 1) sham group, 2) model group, and 3) model group treated with daily gavage administration of 100 mg/kg/d quercetin. Histopathological examination was performed using hematoxylin and eosin (HE) staining. The microstructure of the lesions was examined using electron microscopy. The expressions levels of related proteins, such as the peroxisome proliferator-activated receptor-γ (PPARγ), methionine adenosyl-transferase 2A (MAT2A), Ki67 and VEGF was measured using Western blotting or Immunohistochemistry.

Compared to the model group, the medication group showed sparse endometrial stromal cells, irregular morphology, and numerous vacuoles, indicating apoptosis. Compared to the sham group, SAM expression was unchanged (P > 0.05), while PPARγ decreased. MAT2A, PRMT5, cyclin D1, and C-MYC increased, and vimentin, Ki67, VEGF, and caspase-1 were strongly positive (P < 0.05). Quercetin intervention reduced ectopic lesion weights, increased PPARγ, and decreased MAT2A, PRMT5, SAM, cyclin D1, and C-MYC. Vimentin, Ki67, VEGF, and caspase-1 were weakly positive (P < 0.05).

These results indicate that quercetin effectively reduced endometriosis lesions by modulating key protein expressions and promoting apoptosis.

Quercetin modulated the transcription of the MAT2A/PRMT5 gene by activating PPARγ activity, thereby influencing the ectopic implantation and growth of endometrial cells.

Diet is a modifiable factor that influences several chronic diseases, making lifelong dietary interventions critically important for reducing disease risk. Hence, this study aims to assess the potential causal relationship between diet and sleep apnea (SA).

We analyzed genome-wide association study (GWAS) data from approximately 450,000 individuals, focusing on 8 dietary intakes and GWAS statistics for 249 metabolites from the UK Biobank. Sleep apnea-related phenotypic data from 16,761 participants were sourced from the FinnGen Biobank. Furthermore, we conducted a series of two-sample Mendelian Randomization (two-sample MR) to explore the causality between diet and SA. Sensitivity analyses were conducted to assess the robustness of the two-sample MR results, and reverse MR analysis was performed to examine potential reverse causality. Multivariate MR (MVMR) analysis and mediation effect estimation were employed to evaluate the mediating roles of metabolites.

Two-sample MR analyses revealed significant causal associations between bread intake (OR=0.56, 95% CI 0.35–0.89, P =0.014), cheese intake (OR=0.67, 95% CI 0.50–0.89, P =0.006), and dried fruit intake (OR=0.61, 95% CI 0.39–0.95, P =0.029) with SA. Reverse MR analysis indicated a causal effect of SA on dried fruit intake (P < 0.05). Univariate MR analyses further identified significant causal effects of bread and cheese intakes on 2 and 32 metabolites, respectively (P < 0.05). Subsequent MVMR analysis demonstrated direct causal effects of bread and cheese intake on SA, independent of metabolite mediation (P < 0.05). Furthermore, the mediating effect of cheese intake on SA through glucose was estimated at 0.023 (90% CI 0.01–0.046), whereas other modeled mediation effects were not statistically significant.

The MR analysis in this study offers genetic evidence indicating that heightened genetic susceptibility to cheese and bread intake potentially reduces SA risk. These findings underscore and validate the significance of diet in preventing SA.

Trauma, resulting from mechanical factors, entails damage to human tissues or organs. Whether occurring during times of war or peace, trauma is prevalent, particularly skin defects arising from surgery or external injuries. The development and design of effective wound dressings have become paramount. Bingqing Gao(BQG), rooted in Chinese folk medicine, is employed explicitly in trauma treatment based on traditional Chinese medicine (TCM) theory. This study aims to elucidate how BQG facilitates full-thickness skin wound healing in Sprague Dawley (SD) rats.

Data collection commenced using two approaches: retrieval from TCM system pharmacology databases (TCMSP) and literature mining to compile the practical chemical components and targets of BQG. A drug-target network was constructed. Subsequently, disease targets related to wound healing were collected to select core targets and pathways, building a drug-disease target protein-protein interaction (PPI) network using the ClusterONE algorithm to identify core genes. Gene Ontology (GO) and KEGG enrichment analyses were conducted based on the Metascape database. Finally, molecular docking validation was performed on the screened core targets and core components. In terms of in vivo experimentation, an SD rat full-thickness skin defect model was established, and varying doses of BQG were applied. Healing area, HE staining, Masson staining, ELISA, PCR, and other methods were employed to validate cytokines, differential proteins, and pathways. The study collectively discusses the mechanism and targets by which BQG promotes full-thickness skin wound healing in SD rats.

Through network pharmacology screening, we identified various active components, including resveratrol, Lithospermic acid B, sanguiinH-2, asernestioside A_qt, kaempferol, daidzein, quercetin, apigenin, and Medicarpin. The core targets encompass Interleukin-6 (IL-6), Protein Kinase B (AKT1), Vascular Endothelial Growth Factor A (VEGFA), Interleukin-1 beta (IL-1β), Tumor Protein 53 (TP53), Epidermal Growth Factor Receptor (EGFR), Tumor Necrosis Factor (TNF), Albumin (ALB), among others. Potential signaling pathways include Phosphoinositide 3-kinase (PI3K)/AKT, Tumor Necrosis Factor (TNF), Hypoxia-Inducible Factor-1 (HIF-1), and more. Molecular docking studies suggest a robust binding interaction between the active components of BQG and disease targets, indicating a potential regulation of cytokines through the PI3K/AKTsignaling pathway, thereby promoting wound healing. The results of the in vivo experiment revealed that, in comparison to the model group, both the rhb-FGF group and BQG-H group exhibit a noteworthy increase in the expression levels of PI3K and AKT genes. Concurrently, there is a significant decrease in the levels of pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. Additionally, there is a substantial increase in the levels of Transforming Growth Factor-beta (TGF-β) and Vascular Endothelial Growth Factor (VEGF).

Network pharmacology results indicate that BQG promotes wound healing through multiple components, targets, and pathways. In vivo experimental results suggest that BQG may activate the PI3K/AKTsignaling pathway, inhibit the production and release of related pro-inflammatory cytokines IL-1β, IL-6, and TNF-α, promote VEGF generation at the injury site, and enhance TGF-β signaling transduction, effectively regulates the inflammatory response at the site of injury, promotes vascular regeneration in the injury area, and induces the proliferation and migration of cells in the injury area, ultimately contributing to wound healing. This study establishes the foundation for a more profound understanding of the molecular mechanisms underlying BQG's promotion of wound healing and offers insights for future drug research on BQG.