Cnidii Fructus (CF) is a Chinese herbal medicine with bioactive immune properties and potential applications in treating ulcerative colitis (UC), but its well-defined bioactive constituents are understudied. The current study aimed to investigate CF's anti-UC bioactivity and elucidate its bioactive constituents via chromatographic analysis, in vitro/in vivo tests, and spectrum-effect analysis. Results showed 10 batches of CF inhibited the inflammatory response induced by lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. High-performance liquid chromatography (HPLC) fingerprint profiles of the 10 batches were established, with 16 common peaks identified. Subsequently, 15 of these peaks (including eight reference-validated compounds) were characterized by ultraperformance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS). Four anti-UC-related compounds were identified via spectrum-effect relationships, namely, diosmin, xanthotoxol, imperatorin, and osthole. Experimental verification confirmed these as key components potentially linked to alleviating inflammatory responses in UC. This study is the first to establish a spectrum-effect relationship for CF's anti-UC effects, identify its critical active constituents, and provide a scientific basis for CF quality control and UC therapeutic development.

OBJECTIVES:Ulcerative colitis (UC) is a chronic, relapsing inflammatory bowel disease. Recent research has focused on microbial dysbiosis within the gut environment. Although Shengjiang San modification ameliorates inflammation via activation of the PKC signaling pathway, the specific molecular mechanisms underlying its anti-inflammatory effects remain unclear. This study aimed to systematically investigate the anti-inflammatory mechanisms of Shengjiang San modification. METHODS:UC models were established in 50 male SD rats using Dextran Sulfate Sodium Salt (DSS). Rats were divided into model group, mesalazine group, and Shengjiang San modification high/mid/low-dose groups. After 14 days of intervention, samples were collected. General condition, gross colon morphology, and histopathological changes were observed. Untargeted metabolomics was used to detect serum metabolites, 16S rRNA sequencing was performed to analyze gut microbiota, and combined multi-omics analysis was conducted. RESULTS:Shengjiang San modification significantly improved colonic pathological injury and restored body weight. Metabolomics identified 18 aberrant metabolites, with 16 returning to near-normal levels after treatment, primarily involving the protein kinase C (PKC) signaling pathway. Gut microbiota analysis revealed decreased pathogenic bacteria and significantly increased beneficial bacteria. Notably, the lauric acid-Shigella correlation was disrupted. Compared to the control group, the high-dose group showed superior efficacy in restoring the Firmicutes/Bacteroidetes (F/B) ratio. CONCLUSIONS:Shengjiang San modification exerts anti-inflammatory effects and promotes colonic mucosal repair in UC rats by blocking lauric acid-Shigella synergistic damage via the PKC signaling pathway. The high-dose formulation demonstrated optimal therapeutic efficacy.

IntroductionThis research sought to investigate the therapeutic efficacy and underlying mechanisms of Shengjiang Powder (SJP) in the context of chronic gastritis.MethodsA rat model of gastric mucosal injury was induced by 1-methyl-3-nitroso-1-nitrosoguanidine (MNNG) and a 10% NaCl solution, followed by treatment with either Vitacoenzyme or SJP. Histopathological changes in gastric tissues were evaluated using hematoxylin-eosin staining. Serum IL-6 levels were measured by enzyme-linked immunosorbent assay. Western blot (WB) analysis was performed to detect COX-2 expression. Immunohistochemistry was used to assess TFF1 and TFF2 expression. Serum metabolomics was conducted to identify differential metabolites and enriched pathways, while proteins related to the FOXO signaling pathway were further examined in vivo. WB analysis was used to detect SIRT1, FOXO1, and STAT3. CAT and SOD2 activities were measured using biochemical assays. Multiplex immunohistochemistry was used to examine SIRT1-FOXO1 co-localization and FOXO1 nuclear localization. For the in vitro experiments, normal human gastric epithelial cells were treated with MNNG to induce inflammatory injury, followed by SJP administration in the presence or absence of the SIRT1 inhibitor EX-527.ResultsSJP alleviated gastric mucosal injury, reduced serum IL-6 levels and COX-2 expression, and significantly increased TFF1 and TFF2 expression. Metabolomics analysis identified 13 differential metabolites. The targets of these metabolites were intersected with gastritis-related targets, yielding 522 overlapping targets that were significantly enriched in the FOXO signaling pathway. In vivo and in vitro experiments showed that SJP upregulated SIRT1 and FOXO1 expression, increased CAT and SOD2 activities, inhibited STAT3 expression, and promoted FOXO1 nuclear localization. EX-527 blocked the restorative effects of SJP on FOXO1 expression and antioxidant enzyme activities.ConclusionSJP ameliorates gastric mucosal injury and inflammation, potentially through modulation of the SIRT1/FOXO1 axis.

Cinnamic acid (CA), the primary bioactive component of cinnamon bark, exhibits therapeutic potential against myocardial injury. This study examined CA's cardioprotective effects in a rat myocardial injury model induced by arsenic trioxide (ATO). CA treatment significantly reduced cardiac enzyme levels and pathological changes versus ATO controls. It decreased Reactive Oxygen Species production, upregulated Nuclear factor erythroid 2-related factor 2/Glutathione S-Transferase Mu 2 transcription, enhanced antioxidant activities, and reduced inflammatory markers. Transmission Electron Microscopy demonstrated preserved mitochondrial ultrastructure. Western blot analysis revealed that CA elevated B-cell lymphoma-2 while suppressing pro-apoptotic proteins, and enhanced AMP-activated protein kinase α2 (AMPKα2), phosphorylated AMPKα2 (p-AMPKα2), Sirtuin 1 (SIRT1) and Proliferator-activated receptor-gamma coactivator 1α (PGC-1α) expression. Molecular docking confirmed the robust binding affinity of CA toward pathway-associated proteins. These findings indicate CA alleviates ATO-induced myocardial injury through AMPKα2/SIRT1/PGC-1α pathway modulation, suppressing Reactive Oxygen Species, oxidative stress, inflammation, mitochondrial dysfunction, and apoptosis.

Background:Gastric cancer (GC) is a leading cause of cancer-related mortality worldwide, and lipid metabolism reprogramming plays a crucial role in tumor progression. ATP citrate lyase (ACLY), a key enzyme in de novo lipid synthesis, has been implicated in multiple cancers, but its role in GC remains incompletely understood. This study aimed to investigate the expression, prognostic significance, and potential molecular mechanisms of ACLY in GC, with a particular focus on its relationship with lipid metabolism and ubiquitination. Methods:VOSviewer and Bibliometrix were used for analysis of the PubMed database related to GC and lipid metabolism. The expression levels of ACLY in GC tissues and normal tissues in The Cancer Genome Atlas (TCGA) database were compared and western blot was used to detect. The relationship between ACLY and the prognosis of GC patients was analyzed through bioinformatics. Genome Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) functional enrichment were performed for ACLY-related genes, and three gene data related to ACLY, lipid metabolism and GC were analyzed. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analyzed substances in 2-furoic acid-treated and normal HGC-27 cells. Results:Lipid metabolism and patient prognosis is a hot topic in the field of GC. Compared with that in normal tissues, the level of ACLY in GC tissues is significantly greater, and it is associated with poor patient prognosis. The prognosis of patients with GC is related to age, node (N) stage and metastasis (M) stage. The functions of the ACLY-related genes in cell differentiation and the structure, signal transduction and enzyme activity of GC are enriched. ACLY is correlated with lipid metabolism genes in GC. The content of fatty acids in GC cells decreased after 2-furoic acid treatment. Conclusions:ACLY regulates lipid metabolism to affect the occurrence and development of GC.

Baihe Dihuang Tang (BDT), is a traditional Chinese medicinal formulation historically utilized to manage various health conditions, including insomnia. This therapeutic use of BDT for treating insomnia is rooted in its potential to regulate the gut microbiota, neuroendocrine, and serotonin systems, which may collectively contribute to its effectiveness. This study aims to explore the anti-insomnia effects of BDT, and focus on its underlying mechanisms, emphasizing the potential interplay with gut microbiota, neuroendocrine, and serotonin pathways. An insomnia mouse model was induced using p-chlorophenylalanine (PCPA). Subjects received varying doses of BDT or a saline solution as a control. Behavioral assessment was conducted via the open field test and elevated plus maze test. Hypothalamic monoamine neurotransmitter levels were quantified using ELISA kits. Neurosteroid levels in brain and serum samples were determined through high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). Gut microbiota composition was evaluated using 16S rRNA amplicon sequencing. PCPA-induced insomnia led to significant alterations in neurosteroids, monoamine neurotransmitters, and gut microbiota composition. BDT treatment markedly improved behavioral parameters in insomniac mice, evidenced by enhanced motility and reduced sleep latency compared to controls. BDT administration restored neurosteroid and monoamine neurotransmitter dose-dependently, suggesting potential for neuroendocrine system homeostasis restoration. BDT-treated mice exhibited significant gut microbiota composition changes, including reduced Acidobacteria, increased Fusobacteria and Firmicutes at the phylum level, and decreased Alistipes at the genus level, compared to the insomnia model group. BDT effectively rectifies gut dysbiosis and mitigates neuroendocrine and serotonin system dysfunctions induced by insomnia, emerging as a promising therapeutic candidate for insomnia management.

The transient receptor potential melastatin 7 channel (TRPM7) is a nonselective cation channel highly expressed in some human cancer tissues. TRPM7 is involved in the proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) of cancer cells. Modulation of TRPM7 could be a promising therapeutic strategy for treating cancer; however, efficient and selective pharmacological TRPM7 modulators are lacking. In this study we investigated N- [4- (4, 6-dimethyl- 2-pyrimidinyloxy) - 3- methylphenyl] -N' - [2 -(dimethylamino)] benzoylurea (SUD), a newly synthesized benzoylurea derivative, for its effects on cancer cell migration and EMT and on functional expression of TRPM7. Our previous studies showed that SUD induces cell cycle arrest and apoptosis of MCF-7 and BGC-823 cells (human breast cancer and gastric cancer cell lines, respectively). Here, we show that SUD significantly decreased the migration of both types of cancer cells. Moreover, SUD decreased vimentin expression and increased E-cadherin expression in both cell types, indicating that EMT is also decreased by SUD. Importantly, SUD potentially reduced the TRPM7-like current in a concentration-dependent manner and decreased TRPM7 expression through the PI3K/Akt signaling pathway. Finally, molecular docking simulations were used to investigate potential SUD binding sites on TRPM7. In summary, our research demonstrated that SUD is an effective TRPM7 inhibitor and a potential agent to suppress the metastasis of breast and gastric cancer by inhibiting TRPM7 expression and function.

Chronic atrophic gastritis (CAG) is a chronic inflammatory disease and precancerous lesion in stomach cancer. Abnormal activation cellular ferroptosis further damages gastric tissue, which is susceptible to inflammation. Luteolin has powerful anti-inflammatory and regulatory potential for cellular ferroptosis. We aimed to clarify the involvement of luteolin in inflammation and ferroptosis during CAG. Luteolin targets were searched to identify intersecting genes in the chronic atrophic gastritis disease database. The AGE-RAGE pathway is a potential target of luteolin for the treatment of chronic atrophic gastritis and a binding site between luteolin and RAGE was predicted through a computer simulation of molecular docking. We established a CAG rat model using N-methyl-N-nitro-N-nitroguanidine. The therapeutic effect of luteolin on CAG was detected using western blotting, qPCR, hematoxylin and eosin staining, lipid oxidation (MDA), and Fe2+ assays. Luteolin inhibited the AGE-RAGE signaling pathway and reduced the inflammatory response in gastric tissues. Additionally, luteolin downregulated the concentration of (MDA) and Fe2+, and CAG downregulated the expression levels of ACSL4 and NOX1 and upregulated the expression levels of FIH1 and GPX4 ferroptosis-related proteins, thus inhibiting the ferroptosis of gastric tissue cells, which had a therapeutic effect on CAG.

The aim of this study was to identify the potential compounds, core targets and possible mechanism of Huanglian Jiedu decoction in treating ulcerative colitis based on network pharmacology and molecular docking. At first, potential compounds of Huanglian Jiedu decoction were retrieved from traditional Chinese medicine systems pharmacology. And then, the targets related to compounds and ulcerative colitis was obtained from traditional Chinese medicine systems pharmacology, Online Mendelian Inheritance in Man, GeneCards and DisGeNET. Next, Cytoscape was used to visualize drug-compound-common target-disease network and protein-protein interaction network. Moreover, gene ontology and Kyoto encyclopedia of genes and genomes enrichment analysis was performed by database for annotation, visualization and integrated discovery to investigate possible mechanism of Huanglian Jiedu decoction against ulcerative colitis. At last, molecular docking verified the reliability of the prediction results. 55 compounds and 84 targets of Huanglian Jiedu decoction were screened out as potential players on ulcerative colitis. After network analyses, 10 core compounds (quercetin, kaempferol, wogonin, baicalein, acacetin, 5-hydroxy-7-methoxy-2-(3,4,5- trimethoxyphenyl) chromone, beta-sitosterol, moslosooflavone, 5,7,4'-trihydroxy-8-methoxyflavone, oroxylin A) and 10 core targets (interleukin-6, interleukin-1 beta, tumor necrosis factor-alpha, threonine-protein kinases, tumor antigen p53, prostaglandin-endoperoxide synthase 2, JUN, CXCL8, C-C motif chemokine 2, matrix metalloproteinase-9) were identified. Furthermore, the inflammatory response, tumor necrosis factoralpha signaling pathway, pathways in cancer, T cell receptor signaling pathway, toll-like receptor signaling pathway and nuclear factor kappa B signaling pathway may be involved in the treatment of ulcerative colitis using Huanglian Jiedu decoction. This study reveals that Huanglian Jiedu decoction contains multiple ingredients, multiple targets and multiple pathways in treating ulcerative colitis, which provides a basis for further research.

This study aims to investigate the effect of modified Danggui Shaoyao Powder on the suppressor of cytokine signaling 3(SOCS3)/Toll-like receptor 4(TLR4) signaling pathway in gastric tissue of rats with chronic atrophic gastritis(CAG).Sixty SPF-grade SD rats were randomly assigned into the normal group, model group, Moluo Pills group, and high-, medium-, and low-dose groups of modified Danggui Shaoyao Powder.The rats in other groups except the normal group were treated with N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) to establish the CAG model.After 12 weeks of modeling, the rats in each group were administrated with corresponding drugs by gavage for 8 weeks.After the last administration, the histopathological changes of rat gastric mucosa were observed via hematoxylin-eosin(HE) staining.The serum levels of IL-6, TNF-α, and CRP were determined by enzyme-linked immunosorbent assay(ELISA).The mRNA levels of SOCS3 and TLR4 were determined by real-time PCR.The protein levels of SOCS3, TLR4, JAK2, p-JAK2, STAT3, and p-STAT3 in rat gastric tissue were measured by Western blot.Immunohistochemical method was employed to determine the protein levels of NF-κB, MyD88, NLRP3, Bcl-2, Bax, and Bad in rat gastric tissue.The results showed that modified Danggui Shaoyao Powder alleviated gastric mucosal atrophy of rats, significantly lowered the levels of IL-6, TNF-α, and CRP in rat serum, up-regulated the mRNA level of SOCS3, and down-regulated the mRNA level of TLR4 in rat gastric tissue.Furthermore, modified Danggui Shaoyao Powder up-regulated the protein level of SOCS3, down-regulated the protein levels of TLR4, p-JAK2, p-STAT3, NF-κB, MyD88, NLRP3, Bax, and Bad, and promoted the expression of Bcl-2 protein.Therefore, modified Danggui Shaoyao Powder may mitigate the gastric mucosal atrophy of rats by regulating the SOCS3/TLR4 signaling pathway.

Leptin is the pivotal modulator in the onset and progression of breast cancer and obesity. Celastrol, which is extracted from the roots of Tripterygium wilfordi plants, exerts various anticancer bioactivities and has recently emerged as a candidate to treat obesity by improving leptin sensitivity. However, the relationship between leptin and celastrol in the treatment of breast cancer is unknown. Here, the growth and migration of MCF-7 cells induced by leptin were tested to demonstrate the antineoplastic activity of celastrol. Transcriptomic analysis and western blotting were conducted to explore the biological roles of leptin in treating breast cancer with celastrol. The present findings showed that celastrol remarkably reversed leptin-triggered cell proliferation and migration in MCF-7 cells. Fifty-two mRNAs with fivefold higher counts and 149 mRNAs with fivefold lower counts were identified in the celastrol-treated MCF-7 cells. According to the GO and KEGG analyses, the effects of celastrol on MCF-7 cells forced lipid metabolism and the endocrine system. Moreover, leptin treatment induced phosphorylation of leptin receptor and PI3K/AKT in MCF-7 cells, whereas pretreatment with celastrol partly abrogated leptin activation. The binding of celastrol to the leptin receptor was also confirmed by molecular docking. The antitumor effect of celastrol is proposed to be mediated by its binding to the leptin receptor and controlled downregulation of the PI3K/AKT pathway.

To analyze pesticide residues, GC coupled with quadrupole-Orbitrap MS (GC-Orbitrap-MS) has become a powerful tool because of its unique characteristics of accurate mass full-spectrum acquisition, high resolution, fast acquisition rates, and overcoming matrix interference. This paper presents an efficiency evaluation of GC-Orbitrap-MS for identification and quantitation in the 352 pesticide residues analysis of chrysanthemum flowers in full-scan mode. A streamlined pretreatment approach using one-step extraction and dilution was used, which provided high-throughput processing and excellent recovery. The samples were extracted using acetonitrile. The extracted solution was purified by a Sin-QuEChERS Nano column to suppress the matrix in chrysanthemum flowers and determined by GC-Orbitrap-MS. The calibration curves for the 352 pesticides obtained by GC-Orbitrap-MS were linear in the range of 0.5–200  μ g·kg −1 , with the correlation coefficients higher than 0.99. The limits of detection (LODs) and the limits of quantification (LOQs) for the 352 pesticide residues were 0.3–3.0  μ g·kg −1 and 1.0–10.0  μ g·kg −1 , respectively. The average recoveries in chrysanthemum flower at three levels were 95.2%, 88.6%, and 95.7%, respectively, with relative standard deviations (RSDs) of 7.1%, 7.5%, and 7.2%, respectively. Lastly, the validated method and retrospective analysis was applied to a total of 200 chrysanthemum flower samples bought in local pharmacies. The proposed method can simultaneously detect multipesticide residues with a good performance in qualitative and quantitative detection.

目的 基于网络药理学筛选雷公藤治疗乳腺癌主要活性成分并初步探讨分子机制.方法 利用TCMSP平台获取雷公藤活性成分及作用靶点信息,检索GeneCard数据库筛选雷公藤治疗乳腺癌潜在靶标,将两者进行匹配获得雷公藤治疗乳腺癌核心成分及靶点基因.应用Cytoscape3.6.0、AutoDock Vina等软件,构建活性成分-靶点网络图,进行靶点基因富集分析,完成核心靶标与成分的分子对接.利用CCK8及qPCR等方法验证核心单体成分对乳腺癌细胞及治疗靶点的作用.结果 筛选获得雷公藤治疗乳腺癌有效成分7种,核心治疗靶点25个.其中最优成分为雷公藤红素,其对乳腺癌细胞具有明显抑制作用且显著降低核心治疗靶点的mRNA表达.结论 雷公藤通过多靶点发挥治疗乳腺癌作用,其中雷公藤红素是其核心药用成分.

Background: Claudins (CLDNs) are a family of closely related transmembrane proteins that have been linked to oncogenic transformation and metastasis across a range of cancers, suggesting that they may be valuable diagnostic and/or prognostic biomarkers that can be used to evaluate patient outcomes. However, CLDN expression patterns associated with colorectal cancer (CRC) remain to be defined. Methods: The mRNA levels of 21 different CLDN family genes were assessed across 20 tumor types using the Oncomine database. Correlations between these genes and patient clinical outcomes, immune cell infiltration, clinicopathological staging, lymph node metastasis, and mutational status were analyzed using the GEPIA, UALCAN, Human Protein Atlas, Tumor Immune Estimation Resource, STRING, Genenetwork, cBioportal, and DAVID databases in an effort to clarify the potential functional roles of different CLDN protein in CRC. Molecular docking analyses were used to probe potential interactions between CLDN4 and TGFβ1. Levels of CLDN4 and CLDN11 mRNA expression in clinical CRC patient samples and in the HT29 and HCT116 cell lines were assessed via qPCR. CLDN4 expression levels in these 2 cell lines were additionally assessed following TGFβ1 inhibitor treatment. Results: These analyses revealed that COAD and READ tissues exhibited the upregulation of CLDN1, CLDN2, CLDN3, CLDN4, CLDN7, and CLDN12 as well as the downregulation of CLDN5 and CLDN11 relative to control tissues. Higher CLDN11 and CLDN14 expression as well as lower CLDN23 mRNA levels were associated with poorer overall survival (OS) outcomes. Moreover, CLDN2 and CLDN3 or CLDN11 mRNA levels were significantly associated with lymph node metastatic progression in COAD or READ lower in COAD and READ tissues. A positive correlation between the expression of CLDN11 and predicted macrophage, dendritic cell, and CD4+ T cell infiltration was identified in CRC, with CLDN12 expression further being positively correlated with CD4+ T cell infiltration whereas a negative correlation was observed between such infiltration and the expression of CLDN3 and CLDN15. A positive correlation between CLDN1, CLDN16, and neutrophil infiltration was additionally detected, whereas neutrophil levels were negatively correlated with the expression of CLDN3 and CLDN15. Molecular docking suggested that CLDN4 was able to directly bind via hydrogen bond with TGFβ1. Relative to paracancerous tissues, clinical CRC tumor tissue samples exhibited CLDN4 and CLDN11 upregulation and downregulation, respectively. LY364947 was able to suppress the expression of CLDN4 in both the HT29 and HCT116 cell lines. Conclusion: Together, these results suggest that the expression of different CLDN family genes is closely associated with CRC tumor clinicopathological staging and immune cell infiltration. Moreover, CLDN4 expression is closely associated with TGFβ1 in CRC, suggesting that it and other CLDN family members may represent viable targets for antitumor therapeutic intervention.

Collagen type IV (Col IV) is the main constituent of the basement membrane. Under physiological conditions, Col IV plays an important role in maintaining epithelial integrity and stabilizing epithelial function of the gastric mucosa. Under pathological conditions, Col IV can be free from the basement membrane under the influence of tumor cells and play a pro-metastatic role. Although there is a increasing number of investigations on Col IV, no studies to date have directly uncovered the prognostic role and potential regulatory role of the six isoforms of Col IV in gastric cancer. In the present experiment, we aimed to analyze the role of COL4A family genes in gastric cancer. COL4A1/2/3/4 was significantly overexpressed, while COL4A5/6 was decreased in gastric cancer tissues according to TCGA data and our immunohistochemical staining results. And COL4A1 had a positive correlation with tumor stage, while COL4A5/6 had negative correlations with tumor stage. COL4A1/2/4/5/6 can be considered as a diagnostic indicator of gastric cancer. High levels of COL4A1/2/4/5/6 expression may be predictive of a poor prognosis of gastric cancer. The percentages of genetic alterations in COL4As for stomach cancer varied from 3 to 17% based on the TCGA data (COL4A1, 10%; COL4A2, 8%; COL4A3, 3%; COL4A4, 5%; COL4A5, 17%; COL4A6, 17%). Besides, COL4As may modulate tumor progression by participating in classical cancer pathways: COL4A1/2/3/4/5/6 can activate the EMT process; COL4A2/3/4/6 can inhibit apoptosis and cell cycle; COL4A3/4/5 can activate the PI3K/AKT signaling pathway. This study implied that COL4As have diagnostic and prognostic value for gastric cancer.

Clinical evidence from the practice of traditional Chinese medicine (TCM) has shown that Baihe (BH), Dihuang (DH), and their compatibility, Baihe Dihuang Tang (BDT) can treat insomnia. However, the sedative and hypnotic mechanisms of these therapies have not been established. This study evaluated the sedative-hypnotic mechanisms of BH, DH and BDT in terms of their effects on neurotransmitters, the neuroendocrine system and cation-chloride co-transporters in a p -chlorophenylalanine (PCPA)-induced insomnia mice model. The insomnia mice model was established by intraperitoneal injection of PCPA. Mice were randomly assigned into four groups that were treated with BH, DH and BDT (2.25 g/kg, 2.25 g/kg, 4.5 g/kg) and saline solution (wt., 0.9%). Behavioral analyses showed that, compared to mice in the control group, motion activities of mice with insomnia were significantly reduced, sleep latency was prolonged while sleep duration was shortened. Compared to control mice, 5-HT and KCC2 levels were decreased, whereas CRH and ACTH levels were elevated in the insomniac mice. Treatment with BH, DH and BDT ameliorated the symptoms of insomnia and significantly reversed the abnormal levels of the aforementioned neurotransmitters. Notably, BDT effects were the most obvious. These results indicate that BH, DH and BDT improves insomniac symptoms by modifying neurotransmitters levels, the neuroendocrine system and cation-chloride co-transporters.

The transmembrane member 16A (TMEM16A)-encoded Ca2+-activated Cl- channel (CaCC) is expressed in interstitial cells of Cajal (ICCs) and involved in the generation of the slow-wave currents of gastrointestinal (GI) smooth muscles. TMEM16A modulators have been shown to positively or negatively regulate the contraction of gastrointestinal smooth muscle. Therefore, targeting the pharmacological modulation of TMEM16A may represent a novel treatment approach for gastrointestinal dysfunctions such as constipation and diarrhoea. In this study, evodiamine and rutecarpine were extracted from the traditional Chinese medicine Evodia rutaecarpa and identified as novel TMEM16A inhibitors with comparable inhibitory effects. Their effects on intestinal peristalsis were examined. Whole-cell patch clamp results show that evodiamine and rutecarpine inhibited TMEM16A Cl- currents in CHO cells. The half-maximal inhibition values (IC50) of evodiamine and rutecarpine on TMEM16A Cl- currents were 11.8 ± 1.3 μΜ and 9.2 ± 0.4 μM, and the maximal effect values (Emax) were 95.8 ± 5.1% and 99.1 ± 1.6%, respectively. The Lys384, Thr385, and Met524 in TMEM16A are critical for evodiamine and rutecarpine's inhibitory effects. Further functional studies show that both evodiamine and rutecarpine can significantly suppress the peristalsis in isolated guinea-pig ileum. These findings demonstrate that evodiamine and rutecarpine are new TMEM16A inhibitors and support the regulation effect of TMEM16A modulators on gastrointestinal motility.

Anoctamin-1 (ANO1), also known as transmembrane protein 16A (TMEM16A), is identified as a Ca2+-activated Cl- channel that is expressed in many organs and tissues. It is involved in numerous major physiological functions and especially in tumor growth. By screening 530 natural compounds, we identified cepharanthine as a potent blocker of ANO1 channels with an IC50 of 11.2 ± 0.9 μM and Emax of 92.7 ± 1.7%. The Lys384, Arg535, Thr539, and Glu624 in ANO1 are critical for the inhibitory effect of cepharanthine. Similar to its effect on ANO1, cepharanthine inhibits ANO2, the closest analog of TMEM16A. In contrast, up to 30 μM of cepharanthine showed limited inhibitory effects on recombinant ANO6 and bestrophin-1-encoded Ca2+-activated Cl- currents, but it showed no effects on endogenous volume-regulated anion currents (VRAC). Cepharanthine could also potently suppress endogenous ANO1 currents, significantly inhibit cell proliferation and migration, and induce apoptosis in LA795 lung adenocarcinoma cells. Moreover, animal experiments have shown that cepharanthine can dramatically inhibit the growth of xenograft tumors in mice. The high specificity provided by cepharanthine could be an important foundation for future studies of the physiological role of ANO1 channels, and these findings may reveal a new mechanism of its anticancer effect.

Determining the prognosis of gastric cancer is the most crucial step in the treatment process. Cytokeratins are intermediate filaments found in the intracellular structure of epithelial tissues. Recent researches have focused on determining the relationship between the expression of cytokeratins and the degree and prognosis of tumors. This study aimed to investigate the relationship between the incidence of cytokeratin-20 and cytokeratin-7 in patients with gastric carcinoma with factors influencing the prognosis. In this regard, the study was conducted cross-sectional. The expression of cytokeratin-20 and cytokeratin-7 was evaluated on 50 gastric adenocarcinoma specimens with different degrees of differentiation by the immunohistochemical method. We determined the relationship between the incidence of cytokeratin-20 and cytokeratin-7 with factors affecting the prognosis of patients, including the degree of differentiation of gastric cancer tissue, lymph node involvement, and the depth of tumor invasion. Data were statistically analyzed by Chi-square and Spearman tests. The results showed a statistically inverse relationship between the incidence of cytokeratin-20 and cytokeratin-7 with the degree of tissue differentiation and lymph node involvement in gastric cancer. Although there was a statistically significant relationship between the incidence of tissue invasion in gastric cancer and the incidence of cytokeratin-7, there was no association between the incidence of cytokeratin-20 and tissue invasion. In general, decreased cytokeratin-20 and cytokeratin-7 are associated with decreased tissue differentiation and increased lymph node involvement.