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Background: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have revolutionized the treatment of lung cancer patients with mutated EGFR. However, the efficacy of EGFR-TKIs in wild-type EGFR tumors has been shown to be marginal. Methods that can sensitize EGFR-TKIs to EGFR wild-type NSCLC remain rare. Hence, we determined whether combination treatment can maximize the therapeutic efficacy of EGFR-TKIs. Methods: We established a focused drug screening system to investigate candidates for overcoming the intrinsic resistance of wild-type EGFR NSCLC to EGFR-TKIs. Molecular docking assays and western blotting were used to identify the binding mode and blocking effect of the candidate compounds. Proliferation assays, analyses of drug interactions, colony formation assays, flow cytometry and nude mice xenograft models were used to determine the effects and investigate the molecular mechanism of the combination treatment. Results: Betulinic acid (BA) is effective at targeting EGFR and synergizes with EGFR-TKIs (gefitinib and osimertinib) preferentially against wild-type EGFR. BA showed inhibitory activity due to its interaction with the ATP-binding pocket of EGFR and dramatically enhanced the suppressive effects of EGFR-TKIs by blocking EGFR and modulating the EGFR-ATK-mTOR axis. Mechanistic studies revealed that the combination strategy activated EGFR-induced autophagic cell death and that the EGFR-AKT-mTOR signaling pathway was essential for completing autophagy and cell cycle arrest. Activation of the mTOR pathway or blockade of autophagy by specific chemical agents markedly attenuated the effect of cell cycle arrest. In vivo administration of the combination treatment caused marked tumor regression in the A549 xenografts. Conclusions: BA is a potential wild-type EGFR inhibitor that plays a critical role in sensitizing EGFR-TKI activity. BA combined with an EGFR-TKI effectively suppressed the proliferation and survival of intrinsically resistant lung cancer cells via the inhibition of EGFR as well as the induction of autophagy-related cell death, indicating that BA combined with an EGFR-TKI may be a potential therapeutic strategy for overcoming the primary resistance of wild-type EGFR-positive lung cancers. Highlights: 1. BA is a potent wild-type EGFR inhibitor. 2. BA enhances the anticancer efficacy of erlotinib/gefitinib in NSCLC patients with wild-type EGFR. 3. The combination of BA and gefitinib/osimertinib synergistically blocks EGFR and inhibits the PI3K-AKT-mTOR signaling pathway, leading to autophagic cell death and cell cycle arrest. 4. This combination strategy targets wild-type EGFR in NSCLC and induces autophagy and tumor suppression in vivo. BA, an EGFR inhibitor, sensitizes EGFR-TKIs' activity, indicating that BA plus EGFR-TKIs may be a potential therapeutic strategy to overcome the primary resistance of wild-type EGFR NSCLC. [ABSTRACT FROM AUTHOR]

Phytoalexins play a crucial role in plant immunity. However, the mechanism of how phytoalexin is primed by beneficial microorganisms against broad‐spectrum pathogens remains elusive. This study showed that Bacillus cereus AR156 could trigger ISR against broad‐spectrum disease. RNA‐sequencing and camalexin content assays showed that AR156‐triggered ISR can prime the accumulation of camalexin synthesis and secretion‐related genes. Moreover, it was found that AR156‐triggered ISR elevates camalexin accumulation by increasing the expression of camalexin synthesis genes upon pathogen infection. We observed that the priming of camalexin accumulation by AR156 was abolished in cyp71a13 and pad3 mutants. Further investigations reveal that in the wrky33 mutant, the ability of AR156 to prime camalexin accumulation is abolished, and the mediated ISR against the three pathogens is significantly compromised. Furthermore, PEN3 and PDR12, acting as camalexin transporters, participate in AR156‐induced ISR against broad‐spectrum pathogens differently. In addition, salicylic acid and JA/ET signalling pathways participate in AR156‐primed camalexin synthesis to resist pathogens in different forms depending on the pathogen. In summary, B. cereus AR156 triggers ISR against Botrytis cinerea, Pst DC3000 and Phytophthora capsici by priming camalexin synthesis. Our study provides deeper insights into the significant role of camalexin for AR156‐induced ISR against broad‐spectrum pathogens. Summary statement: Beneficial rhizobacterium AR156 triggered ISR by enhancing camalexin accumulation in Arabidopsis, with WRKY33 acting as a positive regulator. PEN3 and PDR12 are involved in AR156‐induced ISR. Salicylic acid and JA/ET signalling pathways participate in AR156‐primed camalexin synthesis to resist various pathogens. [ABSTRACT FROM AUTHOR]

Clinacanthus nutans (Burm. f.) Lindau, a traditional herb renowned for its anti-tumor, antioxidant, and anti-inflammatory properties, has garnered considerable attention. Although its hepatoprotective effects have been described, there is still limited knowledge of its treatment of acute liver injury (ALI), and its mechanisms remain unclear. To assess the efficacy of Clinacanthus nutans in ALI and to identify the most effective fractions and their underlying mechanism of action. Bioinformatics was employed to explore the underlying anti-hepatic injury mechanisms and active compounds of Clinacanthus nutans. The binding ability of schaftoside, a potential active ingredient in Clinacanthus nutans , to the core target nuclear factor E2-related factor 2 (Nrf2) was further determined by molecular docking. The role of schaftoside in improving histological abnormalities in the liver was observed by H&E and Masson's staining in an ALI model induced by CCl 4. Serum and liver biochemical parameters were measured using AST, ALT and hydroxyproline kits. An Fe2+ kit, transmission electron microscopy, western blotting, RT-qPCR, and DCFH-DA were used to measure whether schaftoside reduces ferroptosis-induced ALI. Subsequently, specific siRNA knockdown of Nrf2 in AML12 cells was performed to further elucidate the mechanism by which schaftoside attenuates ferroptosis-induced ALI. Bioinformatics analysis and molecular docking showed that schaftoside is the principal compound from Clinacanthus nutans. Schaftoside was shown to diminish oxidative stress levels, attenuate liver fibrosis, and forestall ferroptosis. Deeper investigations revealed that schaftoside amplified Nrf2 expression and triggered the Nrf2/GPX4 pathway, thereby reversing mitochondrial aberrations triggered by lipid peroxidation, GPX4 depletion, and ferroptosis. The lead compound schaftoside counters ferroptosis through the Nrf2/GPX4 axis, providing insights into a novel molecular mechanism for treating ALI, thereby presenting an innovative therapeutic strategy for ferroptosis-induced ALI. [Display omitted] • Network pharmacology studies have demonstrated the hepatoprotective effects of schaftoside in Clinacanthus nutans. • Ferroptosis is responsible for CCl 4 -induced acute liver injury. • Schaftoside alleviates CCl 4 -induced ferroptosis in acute liver injury. • Schaftoside inhibits ferroptosis via the Nrf2-GPX4 pathway to protect against acute liver injury. [ABSTRACT FROM AUTHOR]

Objective. The present study is aimed at investigating the biochemical and clinical effects of electroacupuncture in patients with sepsis. Methods. Patients with sepsis treated at Guangdong Provincial Hospital of Chinese Medicine from July 2019 to December 2020 were included. Patients were randomly assigned to treatment with routine Western medicine (WM group) or treatment with Western medicine plus electroacupuncture based on Western medicine (EA group). Indices associated with immune function and clinical efficacy were determined before and at 3 and 5 days after treatment. Indicators of immune function included the percentage of T lymphocyte subsets, natural killer (NK) cells, and soluble programmed death protein 1 (sPD-1) levels. Indicators of clinical efficacy included infection-related indicators in whole blood; levels of tumor necrosis factor-α (TNF-α), C-reactive protein (CRP), and interferon-γ (INF-γ); and assessments using acute physiology and chronic health evaluation-II (APACHE-II) and sequential organ failure assessment (SOFA) scores. Results. Baseline data were not different between WM (N = 30) and EA groups (N = 30). At day 5 following treatment, the level of sPD-1 in the EA group was lower than that in the WM group. Proportions of CD3 + T lymphocytes, CD4 + T lymphocytes, and NK cells, the percentage of lymphocytes, and INF-γ levels in the EA group were significantly higher than those in the WM group. Compared with the WM group, the white blood cell count (WBC), percentage and count of neutrophils, ratio of neutrophils to lymphocytes, and levels of CRP and TNF-α were significantly decreased in the EA group 5 days after treatment. The APACHE-II score of the EA group was significantly lower than that of the WM group 5 days after treatment. Conclusion. Electroacupuncture may regulate the immune function of patients with sepsis through the PD-1 pathway to achieve an anti-inflammatory state and improve clinical symptoms. [ABSTRACT FROM AUTHOR]

Background. Tao-He-Cheng-Qi Formula (THCQF) is a traditional Chinese medicine that has been proven to have antitumor effects. The aim of this study was to elucidate the molecular targets and mechanisms of THCQF against colon cancer and construct a prognostic model based on network pharmacology, bioinformatics analysis, and in vitro experiments. Methods. Potential THCQF compounds and targets were retrieved from the Traditional Chinese Medicine Systems Pharmacology and Bioinformatics Analysis Tool for Molecular Mechanism of Traditional Chinese Medicine databases. Differentially expressed genes for colon cancer were screened in The Cancer Genome Atlas and Gene Expression Omnibus databases. The anticolon cancer mechanisms of THCQF were explored using Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Molecular docking simulations and molecular dynamics analysis were used to evaluate the binding between target proteins and active compounds. Finally, the identified compounds were used to treat colon cancer cells from the HCT116 cell line, and expression of mRNA and protein after relevant posttreatment were tested using real-time polymerase chain reaction and western blotting. Results. A total of 27 anticolon cancer targets of THCQF were selected, among which four genes (CCNB1, CCNA2, IL1A, and MMP3) were shown to effectively predict patient outcomes in a prognostic colon cancer model. GO and KEGG enrichment analyses indicated that the activity against colon cancer of THCQF was associated with the interleukin (IL)-4 and IL-3 signaling pathways. Two compounds in THCQF, aloe emodin (AE) and quercetin (QR), were shown to efficiently bind to cyclin B1, the protein encoded by CCNB1. Finally, incubation of HCT116 cells with AE and QR significantly decreased CCNB1 mRNA expression and cyclin B1 levels. Conclusions. Taken together, the results indicate that AE and QR are the pivotal active compounds of THCQF, and CCNB1 is the main molecular target through which THCQF exerts its anticolon cancer effects. The study findings provide insight for studies investigating the anticancer effects of other traditional Chinese medicines. [ABSTRACT FROM AUTHOR]

Trichosanthes kirilowii Maxim. and Bulbus allii Macrostemi are the components of Gualou Xiebai decoction (GLXB), a commonly used herbal combination for the treatment of coronary heart disease (CHD) in traditional Chinese medicine. Although GLXB is associated with a good clinical effect, its active compounds and mechanism of action remain unclear, which limits its clinical application and the development of novel drugs. In this study, we explored key compounds, targets, and mechanisms of action for GLXB in the treatment of CHD using the network pharmacology approach. We identified 18 compounds and 21 action targets via database screening. Enrichment analysis indicated that the effects of GLXB in patients with CHD are primarily associated with the regulation of signalling pathways for tumour necrosis factor, nuclear factor-kappa B, hypoxia-inducible factor-1, arachidonic acid metabolism, and insulin resistance. GLXB thus exerts anti-inflammatory, antihypoxic, and antiagglutinating effects; regulates lipid metabolism; and combats insulin resistance in CHD via these pathways, respectively. After reverse targeting, we observed that the main active compounds of GLXB in the treatment of CHD were quercetin, naringenin, β-sitosterol, ethyl linolenate, ethyl linoleate, and prostaglandin B1. To explore the potential of these compounds in the treatment of CHD, we verified the affinity of the compounds and targets via molecular docking analysis. Our study provides a bridge for the transformation of natural herbs and molecular compounds into novel drug therapies for CHD. [ABSTRACT FROM AUTHOR]

Radix Astragali and Semen Lepidii (HQ-TLZ) is a commonly used herbal medicine combination for treatment of heart failure, which has a good clinical effect. However, its active components and mechanism of action are not clear, which limits its clinical application and development. In this study, we explored the mechanism of action of HQ-TLZ in the treatment of heart failure based on network pharmacology. We obtained 11 active ingredients and 109 targets from the TCMSP database and SwissTargetPrediction database. Next, we constructed the action network and carried out enrichment analysis. The results showed that HQ-TLZ treatment of heart failure is primarily achieved by regulating the insulin resistance, erbB signaling pathway, PI3K-Akt signaling pathway, and VEGF signaling pathway. After inverse targeting, molecular docking, and literature search, we determined that the equivalent molecular groups of HQ-TLZ in the treatment of heart failure were quercetin and kaempferol. Based on network pharmacology, we reveal the mechanism of action of HQ-TLZ in the treatment of heart failure to a certain extent. At the same time, we determined the composition of the equivalent molecular group. This provides a bridge for the consistency evaluation of natural herbs and molecular compounds, which is beneficial to the development of novel drugs and further research. [ABSTRACT FROM AUTHOR]

Neuroinflammation is one of the main causes of sickness and depressive-like behavior. Rosmarinic acid (RA) has been shown to have a significant anti-neuroinflammatory effect. However, the protective effects and the underlying mechanism of RA on sickness and depressive-like behavior under conditions of neuroinflammation are still unclear. In the present study, we investigated the effects and the underlying mechanism of RA on lipopolysaccharide (LPS)-treated mice with sickness behavior. The behavioral effects of LPS treatment and RA administration were assessed using behavioral tests including a sucrose preference test and an open field test. The neuroprotective effects of RA in conditions of neuroinflammatory injury were determined by HE staining, Nissl staining, and immunofluorescent staining. Moreover, its underlying mechanism was analyzed by using real-time PCR analysis, western blot, and immunofluorescent analysis. The results indicated that RA dramatically mitigated sickness behaviors and histologic brain damage in mice exposed to LPS. In addition, RA administration markedly promoted the expression of brain-derived neurotrophic factor (BDNF)/erythroid 2-related factor 2 (Nrf2), the key regulatory proteins for Nrf2 activation (p21 and p62), the downstream antioxidant enzymes (HO-1, NQO1, GCLC), the autophagy-related proteins (LC3II and Beclin1), and mitochondrial respiratory enzyme genes (ME1, IDH1, 6-PGDH), while reducing the expression of pro-inflammatory genes (CD44, iNOS, TNFα, IL-1β). Moreover, the double-label immunofluorescent analysis revealed that RA increased the fluorescence intensity of LC3 mostly co-localized with neurons and co-expressed with Nrf2. Taken together, our research found that RA could effectively alleviate sickness behaviors and nerve injury caused by neuroinflammation, and its protective effects were mediated by the Nrf2 signaling pathway, which reduced cellular oxidative stress, inflammation, mitochondrial respiratory function damage, and autophagy imbalance. Therefore, RA has the potential to prevent or treat sickness and depressive-like behaviors under conditions of neuroinflammation. • RA was shown to improve sickness and depressive-like behaviors in mice after i.c.v. LPS injection for the first time. • RA was revealed to effectively reduce nerve damage induced by LPS through BDNF/Nrf2 signaling pathway. • RA may prevent neurons damage by inhibiting the imbalance of autophagy in LPS-induced mice. • Evidence was found linking the RA-induced anti-depressive action and inhibiting mitochondrial function damage. [ABSTRACT FROM AUTHOR]

Yes-associated protein (YAP) and transcriptional coactivator with PDZ-binding motif (TAZ) serve as transcriptional co-activators that dynamically shuttle between the cytoplasm and nucleus, resulting in either the suppression or enhancement of their downstream gene expression. Recent emerging evidence demonstrates that YAP/TAZ is strongly implicated in the pathophysiological processes that contribute to cardiovascular diseases (CVDs). In the cardiovascular system, YAP/TAZ is involved in the orchestration of a range of biological processes such as oxidative stress, inflammation, proliferation, and autophagy. Furthermore, YAP/TAZ has been revealed to be closely associated with the initiation and development of various cardiovascular diseases, including atherosclerosis, pulmonary hypertension, myocardial fibrosis, cardiac hypertrophy, and cardiomyopathy. In this review, we delve into recent studies surrounding YAP and TAZ, along with delineating their roles in contributing to the pathogenesis of CVDs with a link to various physiological processes in the cardiovascular system. Additionally, we highlight the current potential drugs targeting YAP/TAZ for CVDs therapy and discuss their challenges for translational application. Overall, this review may offer novel insights for understanding and treating cardiovascular disorders. [Display omitted] [ABSTRACT FROM AUTHOR]

• Activation of sigma-1 receptor ameliorates sepsis-induced myocardial injury. • Activation of sigma-1 receptor attenuates mitochondrial oxidative stress and reduces apoptosis. • Sigma-1 receptor has cardioprotective effects against oxidative stress through the Nrf2/HO1 signaling pathway. Sepsis is a condition that triggers the release of large amounts of reactive oxygen species and inflammatory factors in the body, leading to myocardial injury and cardiovascular dysfunction — an important contributor to the high mortality rate associated with sepsis. Although it has been demonstrated that the sigma-1 receptor (S1R) is essential for preventing oxidative stress, its effectiveness in treating sepsis is yet unknown. This study aimed to investigate the role and mechanisms of S1R activation in sepsis-induced myocardial injury. A model of sepsis-induced myocardial injury was constructed by performing cecum ligation and puncture(CLP) surgery on rats. Flv or BD1047 were intraperitoneally injected into rats for one consecutive week before performing CLP, and then intraperitoneally injected into the rats again 1 h after the surgery.The effects of Flv and BD1047 were detected by HE staining, immunofluorescence staining, IHC staining, echocardiography measurements,TUNEL, oxidative stress detection, TEM, flow cytometry and western blot. We further validated the mechanism in vitro using neonatal rat cardiomyocites and H9C2 cells. S1R protein level was reduced in the hearts of septic rats, whereas administration of Flv, an S1R activator, ameliorated myocardial injury, mitochondrial oxidative stress, and pathological manifestations of sepsis. On the other hand, administration of the S1R inhibitor BD1047 exacerbated the mitochondrial oxidative stress, and apoptosis, as well as symptoms and pathological manifestations of sepsis. In addition, we found that up-regulation of S1R activated the Nrf2/HO1 signaling pathway and promoted nuclear translocation of Nrf2, which activated downstream proteins to generate antioxidant factors, such as HO1, in turn alleviating oxidative stress and countering myocardial damage. By scavenging ROS accumulation and reducing mitochondrial oxidative stress via the Nrf2/HO1 signaling pathway, activation of S1R improves cardiac function, mitigates death of cardiomyocytes, and attenuates sepsis-induced myocardial injury. [ABSTRACT FROM AUTHOR]

• Transcriptome analysis shows the responsive pathways to K + deficiency in grapevines. • Ethylene signal (especially ERFs) are activated after K + deficiency. • Cell wall biosynthesis in roots is inhibited after K + deficiency. • Photosynthesis in shoots is inhibited after K + deficiency. • Ion transporters are induced by K + deficiency. Grapevine is sensitive to potassium (K +) deficiency, while its underlying responsive mechanisms are unknown. In this study, transcriptome profiling of grapevine seedlings under K + deficiency conditions over a time course were investigated. Morphological measurements showed that K + deficiency inhibited the normal growth of grapevine seedlings. RNA-sequencing (RNA-seq) was performed with samples of roots and shoots collected under K + deficiency and normal conditions, and numerous differentially expressed genes (DEGs) induced by K + deficiency were identified, including some ion transporter genes, ethylene-responsive transcription factor (ERF) genes, and cell wall biosynthesis-related genes. In roots, by functional and regulatory pathway analysis, genes in ethylene signal (especially ERF) and in cell wall biosynthesis pathway were upregulated and downregulated, respectively, indicating that ethylene signal was activated, while cell wall biosynthesis was inhibited under K + deficiency treatment. And in shoots, genes in ethylene signal and in photosynthesis pathway were upregulated and downregulated, respectively, revealing that ethylene signal was activated, while photosynthesis was suppressed under K + deficiency treatment. Analysis of transcription factors indicated that ERF family extensively involved in the response to K + deficiency. Taken together, this study reveals the multiple molecular responses, including ethylene signal, ion homeostasis, and other metabolic pathways, to K + deficiency treatment in grapevines. [ABSTRACT FROM AUTHOR]