Abstract Third-generation cutaneous growth factor receptor tyrosine kinase inhibitors have led to impressive advances in the treatment of non-small cell lung cancer. However, acquired resistance significantly limits the clinical application of these agents, with the epidermal growth factor receptor (EGFR)C797S mutation being the primary resistance mechanism. In this study, a series of novel EGFR kinase inhibitors was designed and synthesized for the treatment of the EGFRC797S mutation in non-small cell lung cancer. Structure activity relationship screening revealed that compound 5d was the most effective inhibitor of the C797S mutation. It strongly inhibited Baf3-EGFRL858R/T790M/C797S and Baf3-EGFR 19del/T790M/C797S cell lines, with IC50 values of 0.01836±0.0065 and 0.025±0.005 μM, respectively. Molecular docking studies showed that 5d binds to the EGFRL858R/T790M/C797S and EGFR19del/T790M/C797S proteins effectively. A mechanistic study showed that 5d induced cell cycle arrest in the G2/M phase. Compound 5d caused cell apoptosis in a dose-dependent manner, accompanied by a significant increase in the level of the apoptotic protein cleaved caspase-3. An in vivo xenograft assay of mice with Baf3-EGFR19del/T790M/C797S cells showed that 5d effectively inhibited tumor growth, with inhibition rates of 45.2% (70 mg/kg) and 55.7% (100 mg/kg) being observed. Therefore, 5d is worthy of further investigation as a fourth-generation EGFR inhibitor.

BackgroundIn this study, we aimed to explore the mechanism by which resveratrol promotes cisplatin-induced death of HepG2 cells and to provide a potential strategy for resveratrol in the treatment of cancer.MethodsHepG2 cells were exposed to a range of drug concentrations for 24 h: resveratrol (2.5 μg/mL [10.95 μM], 5 μg/mL [21.91 μM], 10 μg/mL [43.81 μM], 20 μg/mL [87.62 μM], 40 μg/mL [175.25 μM], and 80 μg/mL [350.50 μM]), cisplatin (0.625 μg/mL [2.08 μM], 1.25 μg/mL [4.17 μM], 2.5 μg/mL [8.33 μM], 4.5 μg/mL [15.00 μM], and 10 μg/mL [33.33 μM]), 24 μg/mL (105.15 μM) resveratrol + 9 μg/mL (30.00 μM) cisplatin, and 12 μg/mL (52.57 μM) resveratrol + 4.5 μg/mL (15.00 μM) cisplatin. The interaction of two drugs was evaluated by coefficient of drug interaction (CDI), which was based on the Pharmacological Additivity model. The MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to detect the effect of different concentrations of drugs on cell viability, while transcriptome sequencing was used to identify pathways associated with higher gene enrichment. Synchrotron radiation FTIR microspectroscopy experiments and data analysis were conducted to obtain detailed spectral information. The second-derivative spectra were calculated using the Savitzky–Golay algorithm. Single-cell infrared spectral absorption matrices were constructed to analyze the spectral characteristics of individual cells. The Euclidean distance between cells was calculated to assess their spectral similarity. The cell-to-cell Euclidean distance was computed to evaluate the spatial relationships between cells. The target protein of resveratrol was verified by performing a Western blot analysis.ResultsAfter 24 h of treatment with resveratrol, HepG2 cell growth was inhibited in a dose-dependent manner. Resveratrol promotes cisplatin-induced HepG2 cell death through membrane-related pathways. It also significantly changes the membrane components of HepG2 cells. Additionally, resveratrol changes the morphology of the HepG2 cell membrane by decreasing the expression of PLA2G2.ConclusionResveratrol changes the morphology of the HepG2 cell membrane by decreasing the expression of PLA2G2 and promotes cisplatin-induced HepG2 cell death. The combination of cisplatin and resveratrol can play a synergistic therapeutic effect on HepG2 cells.

Objective: The aim of this research was to probe the effect of isobavachalcone inhibiting the high-fat diet (HFD) and streptozotocin (STZ) induced hyperglycemia and hyperlipemia in mice and its underlying mechanisms.Methods: A mouse model with type 2 diabetes mellitus (T2DM) was established by the HFD and low-dose streptozotocin. T2DM mice were treated with 10 mg/kg of isobavachalcone for 4 weeks. The levels of fasting blood glucose (FBG), blood lipids, and relative proteins in the PI3K/AKT signaling pathway as well as the histological structural changes in the liver were determined.Results: The results showed that the expression levels of FBG (42%, P < .001), triglyceride (70%, P < .001), and glucose transporter 2 were significantly reduced by isobavachalcone. The proteinic and mRNA expressions of IRS1, PI3K, and AKT genes of T2DM mice were dramatically increased with the isobavachalcone treatment. Furthermore, isobavachalcone ameliorated the morphology of the liver in T2DM model mice.Conclusion: The results suggested that isobavachalcone controls the development of T2DM as well as the regulatory mechanism involved in glucose and lipid metabolism through the PI3K/AKT signal pathway in the liver.

目的 通过网络药理学与实验验证相结合的方法探究补骨脂乙素治疗 2 型糖尿病的作用机制.方法 使用ECTM、SwissTargetPrediction、PubChem数据库预测补骨脂乙素的作用靶点,使用 GeneCards数据库预测 2 型糖尿病的潜在靶点.使用Venn数据库进行靶点交互分析,通过STRING数据库计算蛋白相互网络关系(PPI),通过Cytoscape 3.9.1 插件MCODE和Cytohubba筛选核心靶点.通过DAVID数据库进行基因本体(GO)和京都基因与基因组百科全书(KEGG)途径的富集分析.建立胰岛素抵抗的HepG2 细胞模型,通过 2-NBDG检测细胞对葡萄糖摄取的能力,通过RT-PCR和Western blotting检测网络药理学预测的磷脂肌醇-3-激酶(PI3K)/蛋白激酶B(Akt)信号通路相关mRNA及蛋白表达.结果 补骨脂乙素和 2 型糖尿病共同作用的基因 92 个,PPI网络(交互得分≥0.150)包含补骨脂乙素和 2 型糖尿病共同目标的 86 个节点和379 个"边".在MCODE和Cytohubba的筛选结果交叉后,获得了 35 个共同靶点.聚类分析表明,补骨脂乙素作用于 2型糖尿病涉及PI3K/Akt信号通路等多种途径及细胞氧化还原反应等多种细胞进程.补骨脂乙素显著增强了胰岛素抵抗细胞对葡萄糖的摄取能力(P＜0.05、0.01).与模型组相比,补骨脂乙素处理后显著增加了胰岛素受体(INS-R)、胰岛素受体底物1(IRS1)、葡萄糖转运蛋白 2(GLUT2)、PI3K、Akt mRNA的表达(P＜0.01、0.001).与模型组相比,二甲双胍组和补骨脂乙素组的INS-R、IRS1、GLUT2、Akt、PI3K、p-Akt、p-PI3K的蛋白表达显著上调(P＜0.05、0.01、0.001).结论 补骨脂乙素可能通过PI3K/Akt信号通路增加胰岛素抵抗细胞对葡萄糖的摄取能力,发挥其抗 2 型糖尿病作用.

急性胰腺炎是消化系统常见的急腹症,以病因多、进展快为特征,早期诊断及治疗与患者的预后密不可分.在诸多影像学检查中,超声能实时、动态的对胰腺、胆道系统进行全面的扫查,在病因诊断、分级、治疗等方面发挥重要作用.本文就超声在急性胰腺炎的应用现状与前景作一概述,以期为临床急性胰腺炎诊治提供参考.

Purpose: To explore genistein, the most active component of soy isoflavones, on viability, expression of estrogen receptor (ER) subtypes, choline acetyltransferase (ChAT), and glutamate receptor subunits in amyloid peptide 25-35-induced hippocampal neurons, providing valuable data and basic information for neuroprotective effect of genistein in A beta(25-35)-induced neuronal injury. Methods: We established an in vitro model of Alzheimer's disease by exposing primary hippocampal neurons of newborn rats to amyloid peptide 25-35 (20 mu M) for 24 h and observing the effects of genistein (10 mu M, 3 h) on viability, expression of ER subtypes, ChAT, NMDA receptor subunit NR2B and AMPA receptor subunit GluR2 in A beta(25-35)-induced hippocampal neurons. Results: We found that amyloid peptide 25-35 exposure reduced the viability of hippocampal neurons. Meanwhile, amyloid peptide 25-35 exposure decreased the expression of ER subtypes, ChAT and GluR2, and increased the expression of NR2B. Genistein at least partially reversed the effects of amyloid peptide 25-35 in hippocampal neurons. Conclusion: Genistein could increase the expression of ChAT as a consequence of activating estrogen receptor subtypes, modulating the expression of NR2B and GluR2, and thereby ameliorating the status of hippocampal neurons and exerting neuroprotective effects against amyloid peptide 25-35. Our data suggest that genistein might represent a potential cell-targeted therapy which could be a promising approach to treating AD.

This study aimed to investigate the therapeutic effect of curcumin on type 2 diabetes and its underlying mechanisms. A type 2 diabetes mellitus rat model was established by providing high-fat diet and low doses of streptozotocin. Type 2 diabetes mellitus rats were treated with low dose and high dose of curcumin for 8 weeks. The results showed that high-dose curcumin significantly reduced fasting blood glucose, total cholesterol, triglyceride, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, alanine aminotransferase, and aspartate transaminase, liver coefficient, and malondialdehyde levels, and BCL2-Associated X expression in the type 2 diabetes mellitus rats. High-dose curcumin increased the levels of liver superoxide dismutase, catalase, and glutathione; as well as the expression of liver B-cell lymphoma-2, phosphatidylinositol 3-kinase, phosphorylated phosphatidylinositol 3-kinase, protein kinase B, and phosphorylated protein kinase B in type 2 diabetes mellitus rats. Furthermore, it ameliorated the histological structure of the liver and pancreas in diabetes mellitus model rats. However, low-dose curcumin had no significant effect on diabetes mellitus model rats. The results suggest that adequate doses of curcumin controls type 2 diabetes mellitus development as well as the mechanism involved in its anti-apoptotic actions and phosphatidylinositol 3-hydroxy kinase/protein kinase B signal pathway regulation in the liver.

The present study elucidates the possible protective effects of curcumin on beta-cells damaged by oxidative stress and its significance in controlling diabetes mellitus in in vitro experiments. Pancreatic islet (RIN-m5F) cells were treated with 25 mmol/L alloxan (AXN) to induce cell damage and the protective effects of curcumin were observed. The results showed that curcumin significantly promoted the cellular activity of AXN-treated RIN-m5F cells, decreased the ratio of apoptosis, downregulated the level of malondialdehyde, upregulated the levels of superoxide dismutase and reactive oxygen species, increased the expression of Bcl-2, cleaved caspase-3, and cleaved PARP1, and decreased the expression of Bax in AXN-treated cells. These results suggest that curcumin inhibits AXN-induced damage in RIN-m5F cells via antioxidative and antiapoptotic mechanisms.

Curcumin is the main secondary metabolite of Curcuma longa and other Curcuma spp, and has been reported to have some potential in preventing and treating some physiological disorders. This study investigated the effect of curcumin in inhibiting high-fat diet and streptozotocin (STZ)-induced hyperglycemia and hyperlipidemia in rats. Twenty-six male Sprague-Dawley (SD) rats (170-190 g) were randomly divided into a standard food pellet diet group (Control group), a high-fat diet and streptozotocin group (HF + STZ group), and a high-fat diet combined with curcumin and STZ group (HF + Cur + STZ group). Compared with the HF + STZ group, the HF + Cur + STZ group exhibited significantly reduced fasting blood glucose (FBG), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), alanine aminotransferase (AST), and aspartate transaminase (ALT) levels, as well as liver coefficients. In the livers of these rats, the expression of malondialdehyde (MDA) and Bax was downregulated, whereas that of superoxide dismutase (SOD) and Bcl-2 was upregulated. Moreover, the liver histology of these rats was improved and resembled that of the control rats. These results suggest that curcumin prevents high-fat diet and STZ-induced hyperglycemia and hyperlipidemia, mainly via anti-oxidant and anti-apoptotic mechanisms in the liver.

Metformin, a first-line drug for type-2 diabetes, plays a potentially protective role in preventing Alzheimer's disease (AD), but its underlying mechanism is unclear. In this study, Aβ25-35 -treated SH-SY5Y cells were used as a cell model of AD to investigate the neuroprotective effect of metformin, as well as its underlying mechanisms. We found that metformin decreased the cell apoptosis rate and death, ratio of Bcl-2/Bax, and expression of NR2A and NR2B, and increased the expression of LC3 in Aβ25-35 -treated SH-SY5Y cells. Metformin also reduced intracellular and extracellular Glu concentrations, as well as the intracellular concentration of Ca2+ and ROS in Aβ25-35 -treated SH-SY5Y cells. These findings suggest that metformin inhibits Aβ25-35 -treated SH-SY5Y cell death by inhibiting apoptosis, decreasing intracellular Ca2+ and ROS by reducing neurotoxicity of excitatory amino acids, and by possibly reversing autophagy disorder via regulating autophagy process.

We established a model of Alzheimer’s disease in vitro by exposing primary hippocampal neurons of neonatal Wistar rats to the β-Amyloid peptide fragment 25–35, Aβ25–35. We then observed the effects of genistein, a type of soybean isoflavone, on Aβ25–35-incubated hippocampal neuron viability, and the electrophysiological properties of voltage-gated sodium channels (NaV) and potassium channels (KV) in the hippocampal neurons. Aβ25–35 exposure reduced the viability of hippocampal neurons, decreased the peak amplitude of voltage-activated sodium channel currents (INa), and significantly reduced INa at different membrane potentials. Moreover, Aβ25–35 shifted the activation curve toward depolarization, shifted the inactivation curve toward hyperpolarization, and increased the time constant of recovery from inactivation. Aβ25–35 exposure significantly shifted the inactivation curve of transient outward K+ currents (IA) toward hyperpolarization and increased its time constant of recovery from inactivation. In addition, Aβ25–35 significantly decreased the peak density of outward-delayed rectifier potassium channel currents (IDR) and significantly reduced IDR value at different membrane potentials. We found that genistein partially reversed the decrease in hippocampal neuron viability, and the alterations in electrophysiological properties of NaV and KV induced by Aβ25–35. Our results suggest that genistein could inhibit Aβ25–35-induced neuronal damage with changes in the electrophysiological properties of NaV and KV.