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Objective: To investigate the expression of eight subunits of chaperonin containing t-complex 1 (CCT) in thyroid cancer (TC) tissues and their correlation with TC staging, patient prognosis, immune cell infiltration, immune checkpoint expression, and chemotherapy drug sensitivity. Methods: Data from The Cancer Genome Atlas (TCGA) database were used to analyze the expression of each CCT subunit in TC tissues and para-cancerous tissues as well as their relationship with the prognosis of TC patients. The biological function of each CCT subunit was analyzed by the Gene Set Enrichment Analysis (GSEA). Data from the TCGA and TIMER2.0 databases were used to analyze the correlations between the expression of each CCT subunit and the tumor microenvironment, infiltration of immune cells, chemotherapeutic drug sensitivity, and immune checkpoint expression. Results: Database analysis showed high mRNA expression of CCT3, CCT7, and CCT8 and low mRNA expression of CCT1, CCT2, CCT5, and CCT6B in TC tissues (P < 0.01 or P < 0.001). CCT3, CCT6B, and CCT8 mRNA expression were correlated with T staging (P < 0.05 or P < 0.01); CCT6B mRNA expression was associated with lymph node metastasis (P < 0.01); CCT5 mRNA expression was associated with distant metastasis (P < 0.05); and CCT6B may serve as an independent prognostic biomarker for overall survival (OS) in TC patients. CCT subunit expression was primarily enriched in signaling pathways such as graft rejection, complement, and interferongamma.Low expression groups of CCT subunits demonstrated significantly higher TC stromal score, immune infiltration score and composite score than the high expression groups (P < 0.05 or P < 0.01 or P < 0.001), displaying a negative correlation between the expression of each CCT subunit and TC stromal, immune infiltration and composite scores (all P < 0.01). The mRNA expression of each CCT subunit was positively correlated with CD8+ T cell and macrophage infiltration (all P < 0.05), the mRNA expression of most CCT subunits (except CCT6B and CCT7) was positively correlated with neutrophil infiltration (P < 0.05 or P < 0.01 or P < 0.001), whereas the mRNA expression of CCT3, 4, 7, and 8 was negatively correlated with CD4+ T-cell infiltration (all P < 0.05). Compared to the low-expression group, patients with high mRNA expression of most CCT subunit had significantly higher IC50 for chemotherapeutic agents such as sorafenib, levatinib, darafenib, trametinib, vandetanib, and cabozantinib (all P < 0.05 or P < 0.01 or P < 0.001). CCT subunit expression in TC tissues was significantly correlated with the expression of PD-1, PD-L1, PD-L2, CTLA4, CD80, and CD86 (P < 0.05 or P < 0.01 or P < 0.001). Conclusion: CCT complexes may promote TC development by affecting the tumor microenvironment thus impacting patient prognosis, potentially serving as a target for the diagnosis and immunotherapy of refractory TC. [ABSTRACT FROM AUTHOR]

Objective: To engineer a BCMA mutant resistant to γ-secretase cleavage in order to stabilize wild-type BCMA expression after γ-secretase cleavage and to generate target cells for measuring BCMA CAR-T cell cytotoxicity. Methods: Our study aimed to engineer a mutant BCMA protein (BCMA-CD8α TM) that could resist γ-secretase cleavage by replacing the transmembrane domain of the wild-type BCMA protein with the human CD8α sequence. Four different types of cells in which this mutant gene was expressed excessively were engineered, including U266 (U266BCMA Mut), K562 (K562BCMA Mut), SKOV3 (SKOV3BCMA Mut), and CHO (CHOBCMA Mut) cells. BCMA CAR Jurkat cells, loaded with the NFAT-EGFP reporter gene (BCMA-CAR-Jurkat-Reporter), were engineered and cocultured with U266BCMA Mut cells. The expression level of EGFP was detected by FCM in order to indicate the activation level of NFAT. The cytotoxicity of BCMA CAR-T cells against Luciferase-labeled K562BCMA Mut cells was detected by the luciferase assay. Additionally, real-time cell analysis (RTCA) technique was employed to detect the cytotoxicity of BCMA CAR-T cells against SKOV3BCMA Mut and CHOBCMA Mut cells. Results: Application of γ-secretase inhibitor LY411575 to inhibit γ-secretase activity significantly enhanced the expression level of BCMA on the surface of wild-type U266 cells, and the average fluorescence intensity was increased by more than 10 times. However, the expression level of BCMA gradually decreased after removal of inhibitors (P<0.01). BCMA-CD8α TM mutant could resist the cleavage of γ-secretase and expressed stably on the surface of U266 cells (P>0.05). U266 cells and U266 cells overexpressing BCMA-CD8α TM were co-incubated with BCMA-CAR-Jurkat-Reporter cells, both of which could activate the Reporter system and enhance the expression of EGFP, but the effect was more significant in U266 cells overexpressing BCMA-CD8α TM (P<0.01). BCMA-CD8α TM mutants were successfully overexpressed in 3 BCMA-negative target cells, namely K562, SKOV3 and CHO cells, and the expression level of the mutant was only slightly increased under LY411575 treatment. Luciferase assay results showed that under different target-effect ratios BCMA CAR-T cells could all be specific and efficient in killing K562 cells overexpressing BCMA-CD8α TM. RTCA results showed that under different target ratios BCMA CAR-T cells could all effectively recognize and kill SKOV3 and CHO cells overexpressing BCMA-CD8α TM, but Mock-T cells with the same target ratio had no such effect. Conclusion: The BCMA-CD8α TM mutant engineered in this study can resist γ-secretase cleavage and exhibits stable surface expression on various target cells, thus offering various methods for evaluating the efficacy and specificity of BCMA CAR-T cell cytotoxicity in vitro. [ABSTRACT FROM AUTHOR]

Objective: To investigate the effects of berberine (BBR) combined with XAV939 on the migration and apoptosis of osteosarcoma MG-63 cells and its possible mechanisms. Methods: MG-63 cells were cultured, and 20~120 µmol/L of BBR and 5~60 µmol/L of XAV939 were added, respectively. The cytotoxicity of BBR and XAV939 was determined by CCK-8 assay, and the combined index of the two was clarified using Chou-Talalay analysis to determine the intervention dose for the subsequent experiments. MG-63 cells were randomly divided into the blank control (NC) group, the BBR group, the XAV939 group, and the BBR+XAV939 combined group. The effects of BBR and XAV939 treatment alone or in combination on the migratory ability of MG-63 cells were detected by the scratch healing assay and the Transwell assay. Hoechst 33258 staining, JC-1 staining and Annexin V-FITC/PI double staining flow cytometry were used to detect the effects on cell mitochondrial membrane potential and apoptosis. Immunofluorescence was used to datect the expression of BAX protein, WB assay was used to detect the effects on the expression of MMP-2 and BAX in the cells, and qPCR was used to detect the effects on the expression of the MMP-2 gene. Results: BBR and XAV939 inhibited the proliferation of MG-63 cells in a dose-dependent manner, and 30 µmol/L BBR, 7.5 µmol/L XAV939 were selected for subsequent experiments. Compared with the NC group, 24 h after the cells were treated with BBR (30 µmol/L), XAV939 (7.5 µmol/L) and BBR+ XAV939 combination, the migration ability of MG-63 cells was significantly reduced; apoptotic cells were significantly increased (all P <0.05); the mitochondrial membrane potential was decreased (P<0.01); the gene expression of MMP-2, and MMP-2, BAX protein levels were all reduced (all P<0.05). In addition, the effect of the combination group was significantly stronger than that of the monotherapy group (P<0.05 or P<0.01). Conclusion: BBR and XAV939 alone or in combination can inhibit the migration and promote the apoptosis of MG-63 cells. The mechanism may be related to the decreased expression of migration-related protein MMP-2 and increased level of apoptosis-related protein BAX. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

[Objectives]Gram-negative resistant bacteria producing New Delhi metallo-β-lactamase-5(NDM-5)pose great threat to the health of human and domestic animals. The study aimed to explore the inhibitory action and the involved mechanism of allicin on NDM-5, which could provide theoretical support for the development of novel β-lactamase inhibitors. [Methods]The binding site of allicin and NDM-5 was predicted by molecular docking with AutoDock. Mutant vector pET21a-NDM-5-C208A was constructed and transferred into E.coli BL21(DE3)competent cell after sequencing. Sensitive test was performed to verify the phenotype of mutant strain. Checkerboard method and time-kill curves assay were performed to assess the combination effect of allicin and meropenem against NDM-5 mutant strain. Enzyme activity assay was established to compare the inhibitory effect of allicin on the activities of C208 mutant NDM-5 and wild-type NDM-5 by expressing the mutant protein NDM-5-C208A in vitro. [Results]The C208A mutant strain BL21(DE3)pET21a-NDM-5-C208A was successfully constructed confirmed by sequencing. The C208A mutant strain regained susceptibility to cephalosporins and carbapenems, and the combination of allicin and meropenem showed independent effect, but not synergistic effect(FICI increased from 0.375 to 2). Enzyme activity assay showed that C208A mutation of NDM-5 caused the decrease of enzyme activity by 85% in vitro and the mutation resulted in the loss of inhibitory effect of allicin on NDM-5. [Conclusions]The Cys208 is a key site for maintaining the enzymatic hydrolysis activity of NDM-5, and allicin can exert an inhibitory effect on the enzyme activity by binding to this site. The results provide a strong support for developing and applying of allicin as a novel β-lactamase inhibitor. [ABSTRACT FROM AUTHOR]

Fatty acid synthase (FASN) is a key enzyme in the fatty acid synthesis that catalyzes the de novo synthesis of lipids. Most of the fatty acids are of food origin in normal tissues, while fatty acids are dependent on the de novo synthesis of lipids in endometrial cancer cells. Previous studies have shown that FASN expression is associated with the development, progression and poor prognosis of endometrial malignancies. The overexpression of FASN is correlated with mitogen extracellular kinase (MEK)/ extracellular signal -regulated kinase (ERK)/ mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/ protein kinase B (Akt)/ mammalian target of rapamycin (mTOR) signaling pathways. Specific blockade of its activity can significantly inhibit the growth and proliferation of tumor cells. We review the action of FASN and mechanism in endometrial malignancies and the progress of FASN inhibitors such as C75, C93, orlistat and so on, in endometrial cancer or other tumors, with the aim of providing new ideas for the treatment of endometrial tumors [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In order to clarify the resistance status and resistance mechanism of the field population of the small brown planthopper (SBPH), Laodelphax striatellus, to commonly used insecticides, we monitored the resistance level of SBPH collected from Yizheng and Yancheng in Jiangsu Province, Changxing in Zhejiang Province and Lujiang in Anhui Province, to eight insecticides during 2021 to 2022 using rice seedling dipping method. The synergistic effects of three detoxification enzyme inhibitors, piperonyl butoxide (PBO), diethylmaleate (DEM), and triphenyl phosphate (TPP), to buprofezin on the control of SBPH were measured. The results showed that four regions populations of SBPH have developed medium to high level resistance to buprofezin (RR = 61.5-148.8), and moderate level resistance to chlorpyrifos (RR = 14.9-28.3). Furthermore, the four regions SBPH populations exhibited sensitive or low-level resistance to nitenpyram (RR = 0.7-9.9), thiamethoxam (RR = 1.6-8.3), dinotefuran (RR = 2.9-10.0) and sulfoxaflor (RR = 2.5-8.7). In addition, all field populations of SBPH were susceptible to pymetrozine (RR = 1.0-5.0) and triflumezopyrim (RR = 0.5-2.3). PBO, TPP, and DEM displayed no synergism effect on the sensitivity of SBPH to buprofezin, which indicated that these three detoxifying enzymes might play little role in the buprofezin resistance. The results of this study provided a scientific basis for the resistance management and field control of SBPH. [ABSTRACT FROM AUTHOR]

Background and objective Acquired and primary resistance to epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) is still the bottleneck of clinical treatment of advanced non-small cell lung cancer (NSCLC). STE029 is a novel anticancer drug which consists of 3-hydroxy-3-methylglutarylcoenzyme A reductase (HMGCR) inhibitor and novel cancer cell membrane targeting molecular. This study aimed to investigate the reversal mechanism of EGFR-TKI resistance by STE029 in lung adenocarcinoma. Methods CCK8 test was used to test the cell viability and survival rate of EGFR mutated PC9 cell (Gefitinib sensitive), PC9/BB4 cell (acquired Gefitinib resistant), and EGFR wild type A549 cell after treatment of STE029, Gefitinib or combination of both. EdU test was applied to detect changes in cell cycle and Hoechst 33258 was applied to detect apoptosis rate in overcoming the EGFR-TKI resistance. The activity of EGFR/PI3K/Akt, cell cycle and apoptosis signal pathways were examined. In vivo, nude mice were exposed to STE029, Gefitinib and STE029+Gefitinib for 5 wk. And the the tumor volume was measured and tumor weight was obtained on the last day. Results (1) PC9 cells was highly sensitive to Gefitinib, while PC9/BB4 and A549 cell showed significant resistance to Gefitinib treatment; (2) STE029+Gefitinib treatment could significantly decrease the 50% inhibitory concentrarion (IC50) of Gefitinib in PC9, PC9/BB4 and A549 cells (P<0.05, respectively); (3) In PC9 and PC9/BB4 cells, STE029+Gefitinib can block cell cycle and inhibit cell proliferation (P<0.001), while there was no significant difference in apoptosis rate among three drug intervention groups (P>0.05); However, apoptosis rate was increased in STE029+Gefitinib group in A549 cell (P<0.01), while no significance detected in cell proliferation (P>0.05). (4) In PC9 and PC9/BB4 cells, the combination of STE029 and Gefitinib could downregulate p-EGFR, p-Akt, p-Cyclin D1 and Cyclin D1 (P<0.001), and upregulate the expression of GSK-3β (P<0.001), and the expression of cleaved caspase-8, caspase-8 cleaved caspase-9, caspase-9 showed no difference among groups (P>0.05). In A549 cells, the combination of STE029 and Gefitinib could downregulate p-Akt (P<0.001) and upregulate cleaved caspase-8 and cleaved caspase-9 (P<0.001); (5)In vivo, the combination of STE029 and Gefitinib effectively inhibited tumor development and progression compared to STE029 alone or Gefitinib alone, with significant difference (P<0.05) in PC9 and PC9/BB4 xenografted tumor. Conclusion STE029 could sensitize Gefitinib by inhibiting EGFR/PI3K/Akt pathway, blocking the tumor cell cycle and proliferation and inducing apoptosis through caspase-8 and caspase-9 dependent pathway. STE029 deserves further investigations in overcoming EGFR-TKI resistance in lung cancer. [ABSTRACT FROM AUTHOR]

In recent years, inhibitors targeting 4-hydroxyphenylpyruvate dioxygenase (HPPD) have become the hot research in the field of herbicide due to their high activity and low risk of resistance. Quinazoline-2,4-dione has been proved to be a potent scaffold. To further exploit its advantage, 30 novel pyrazole-quinazoline-2,4-dione derivatives were designed and synthesized based on the previous work and existing structure-activity relationship, all the structure of which were determined by HRMS, ¹H NMR and 13C NMR. Furthermore, the in vitro and in vivo activity of the synthesized compounds were evaluated. The result indicated that most of the synthesized compounds exhibited comparable or even better enzyme inhibitory activity than that of the positive control benquitrione. The greenhouse herbicidal activity experiments revealed that the tested compounds showed growth inhibition against the six tested weeds, especially for compound 9-28, which achieved more than 80% inhibition against the six species of target weeds at the dosage of 150 g/hm² and 100% inhibition against Echinochloa crus-galli and Digitaria sanguinalis. Furthermore, the AtHPPD-9-28 complex was obtained, which explained the binding mode between the inhibitor and target enzyme, and brought a novel thought to facilitate a detailed understanding for the discovery of new HPPD inhibitors with improved performance. [ABSTRACT FROM AUTHOR]

Dermatomyositis (DM) is characterized by skin lesions, skeletal muscle inflammation, and involvement of other organs. The pathogenesis of DM includes susceptibility genes, environmental stressors, immune and non - immune inducing mechanisms. DM usually presents as subacute progressive proximal muscle weakness, skin rash, or both. Other common extramuscular findings include various heart abnormalities, interstitial lung disease (ILD), and malignant tumors. The auxiliary examination of DM includes the examination of myositis autoantibodies, myoenzymes and electromyography (EMG). Skin treatments for DM include photosensitivity inhibition drugs and antimalarial drugs. If the DM is associated with muscle involvement, systemic corticosteroids is the first line of treatment, and immunosuppressants are used for refractory diseases or as corticosteroids sparing agents in the event of side effects. Other treatments include oral calcineurin inhibitors, cyclophosphamide, biologic agents and Janus kinase (JAK) inhibitor. [ABSTRACT FROM AUTHOR]

Copyright of Chinese Journal of Cancer Biotherapy is the property of Editorial Office of Chinese Journal of Cancer Biotherapy and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Melanoma is the primary malignant tumor of the skin with a very high morbidity and fatality, of which 50% are caused by viral infection of proto-oncogene BRAF mutation. The combination of BRAF and MEK inhibitors is the main treatment of BRAF mutant melanoma, but there have been some drug resistance and drug side effects. Currently, resistance of BRAF mutant melanoma is considered to include primary resistance, adaptive resistance and secondary resistance, and corresponding molecular mechanisms are related and different. At the same time, in order to reduce the incidence of drug resistance in BRAF mutant melanoma, some progress has been made in basic research and clinical application of new target inhibitors, immunotherapy and epigenetics. In this paper, we will summarize the possible mechanism of drug resistance in BRAF mutant melanoma and the current situation and prospects of optimized treatments. [ABSTRACT FROM AUTHOR]

Objective: To study the effect of lipid storage level in cellular lipid droplets (LDs) on the development of metabolic diseases such as obesity and diabetes. Methods: We built up a high content LD imaging and quantitative analysis system to characterize cellular LDs as three-dimensional dynamic parameters such as the total volume of cellular LDs in a cell, the number of total LDs in a cell, and the average volume of LDs. We selected four types of cells including HeLa, AML-12, COS-7, and 3T3-L1 pre-adipocytes as our cell models. Next, we characterized the various phenotypes of cellular LDs in the four types of cells including LD number and LD volume, under different treatments by oleic acids, gene knockdown using RNA interference, and inhibitors for enzymes. Results: After treatment of cells by using oleic acids, more LDs but not larger LDs were produced in AML-12 cells, while the other three types of cells exhibited more and larger cellular LDs as the concentration of oleic acids increased. After knocking down the transcriptional expressions of 19 critical genes in a neutral lipid synthesis pathway in HeLa cells by using RNA interference, we found that double knockdown of DGAT1 and DGAT2 decreased lipid storage in cellular LDs. However, in COS-7 cells, only DGAT1 knockdown was sufficient to reduce the lipid storage of cellular LDs. Interestingly, after treatment of cells by using DGAT1 and DGAT2 inhibitors, we further found that the four types of cells were classified into two groups, HeLa and AML-12 versus COS-7 and 3T3-L1, based on the different resultant phenotypes of LDs. We measured and confirmed that the two different treated responses of cells resulted from the different transcriptional profiles of DGAT1 and DGAT2. Conclusions: In this study, we built up a high content LD imaging and quantitative analysis system to characterize the number and size of cellular LDs in the four types of cell lines. We found that the various phenotypes of LDs in the four types of cells and these results uncover a relationship between the lipid storage mode of cellular LDs and the transcriptional profiles of enzymes in the neutral lipid synthesis pathway. [ABSTRACT FROM AUTHOR]

Ribonucleotide reductase (RR) regulates dNTP production, catalyzing the reduction of ribonucleotide into deoxyribonucleotide diphosphate, which is essential in DNA synthesis and cell proliferation. RR overexpression is related with the tumorigenesis and tumor progression. At present, RR inhibitors have been used on clinical cancer treatment, and new anticancer drugs have been developed targeting RR and its subunits, which may provide new options for cancer therapy. This review discusses the structure and function of RR, the role of RR on cancer therapy, and future application of RR inhibitors.

Immunotherapy is one of the main strategies of anti-tumor therapy at present, in which immune checkpoint inhibitors (ICIs) are the most widely used drug. ICIs resistance is mediated by a variety of cytokines and immune cells, and the mechanism is complex, which is the main reason for the failure of immunotherapy in cancer patients. Histone deacetylase inhibitor (HDACi), as a class of epigenetic regulatory drugs, plays an important role in regulating cell cycle, proliferation, differentiation, and activity. In recent years, Studies have found that HDACi can not only regulate cell biological characteristics, but also closely related to the improvement of tumor ICIs drug resistance. Therefore, the study on how HDACi enhances the efficacy of ICIs is of great significance to tumor immunotherapy. This article will review the research progress of HDACi combined with ICIs in treating malignant tumors and their related mechanism. [ABSTRACT FROM AUTHOR]

Small cell lung cancer (SCLC), characterized by early metastasis, relapse, relapse and resistance and poor prognosis, still faces difficulties in treatment. Recently, Immunotherapy is a novel treatment for SCLC, researchers are also eager to achieve a breakthrough in targeted treatment of SCLC. Genomic instability of SCLC and sensitivity to cytotoxic chemotherapy, therefore, poly ADP-ribose polymerase (PARP) inhibitors targeting DNA repair related pathways have become a hotspot in the research of SCLC targeted therapy. Studies on PARP inhibitors in SCLC have been conducted in combination with other therapeutic strategies, including the treatment of recurrent SCLC and first-line treatment, as well as maintenance treatment after induction. These studies also explored the predictive markers of PARP inhibitors in SCLC. Although the current results of PARP inhibitors in SCLC are limited, and the predictive markers are also inconsistent, we also see that PARP inhibitors could be a breakthrough for precision medicine of SCLC. [ABSTRACT FROM AUTHOR]

The resistant level, mechainism of resistance of Alopecurus myosuroides against fenoxaprop- P-ethyl were determined. Results of the whole-plant dose-response test showed that population HN-05 from Henan Province had evolved high-level resistance against fenoxaprop-P-ethyl (52.2-fold). Using the gene sequencing method, two amino acid mutations at codon 2041 (Ile-Asn) and 2096 (Gly-Ala) were detected in the ACCase gene of the resistant population HN-05. Pre-treatment with the known metabolic inhibitor, piperonyl butoxide (PBO), enhanced the activity of fenoxaprop-P-ethyl (161.3- fold). So that the GR50 value (active ingredient) of fenoxaprop-P-ethyl to HN-05 is 5.4 g/hm2 . This study confirmed the resistance of A. myosuroides against the ACCase inhibitor, fenoxaprop-P-ethyl. The resistance mechanism were conferred by specific ACCase point mutations at amino acid position Ile-2041-Asn and Gly-2096-Ala and P450s-based metabolism. [ABSTRACT FROM AUTHOR]

Coronaviruses pose significant threats to animal and human health, leading to the development of various infectious diseases. It is critical to develop effective vaccines and antiviral medicines to prevent and treat these diseases. The coronavirus genome encodes several types of proteins, including structural, nonstructural, and accessory proteins. Among them, nonstructural protein 13 (NSP13) helicase plays a crucial role in regulating viral replication and the innate immune response of the host. Therefore, it serves as a vital target for the development of anti-coronavirus drugs. This paper presents a comprehensive review of NSP13 research, covering its source, structure, sequence conservation, unwinding mechanism, enzyme inhibitors, protein interaction, and immune regulation. Additionally, the paper analyzes the current challenges in NSP13 research and aims to provide a theoretical foundation for the development of broad-spectrum antiviral drugs targeting NSP13.

Copyright of Shipin Kexue/ Food Science is the property of Food Science Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Background and objective Lung cancer is one of the common malignant tumors that impair human health. With the development of epigenetics, the researchers found that enhancer of Zeste homolog 2 (EZH2) is highly expressed in lung cancer tissue and its expression is closely related to the prognosis. EZH2 inhibitor can also enhance the sensitivity of tumor cells to a variety of anti-tumor drugs. The purpose of this study is to investigate the effect of combination of EZH2 inhibitor and gefitinib on the proliferation, apoptosis and migration of Gefitinib-resistant lung cancer cells. Methods PC9 and PC9/AB2 cells were used for this study. CCK-8 and EdU experiment were used to detect combined treatment on cell viability and proliferation activity; Wound healing assay and Transwell chamber experiment were used to determine the effects of combination therapy on cell migration ability; Flow cytometry was used to detect the effect of combination therapy on EZH2 and apoptosis; Western blot was used to observe the effect of combination therapy on epidermal growth factor receptor (EGFR) signaling pathway-related proteins expression. Results In gefitinib-resistant cell line PC9/AB2, gefitinib combined with EZH2 inhibitor GSK343 can significantly inhibit cell viability, reduce cell migration and increase cell apoptosis. At the same time, combination therapy can significantly inhibit the expression of EZH2 and phosphorylation EGFR proteins. Conclusion The combination of EZH2 inhibitor GSK343 and gefitinib sensitize PC9/AB2 cell to gefitinib response. This study also suggests that synergistic therapy plays a role in the reversal of EGFR-tyrosine kinase inhibitor (EGFR-TKIs) resistance in lung cancer. [ABSTRACT FROM AUTHOR]

Enzymes are closely associated with the onset and progression of numerous diseases, making enzymes a primary target in innovative drug development. However, the challenge remains in identifying compounds that exhibit potent inhibitory effects on the target enzymes. With the continuous expansion of the total number of natural products and increasing difficulty in isolating and enriching new compounds, traditional high-throughput screening methods are finding it increasingly challenging to meet the demands of new drug development. Virtual screening, characterized by its high efficiency and low cost, has gradually become an indispensable technology in drug development. It represents a prominent example of the integration of artificial intelligence with biopharmaceuticals and is an inevitable trend in the rapid development of innovative drug screening in the future. Therefore, this article primarily focused on systematically reviewing the recent applications of virtual screening technology in the development of enzyme inhibitors and explored the prospects and advantages of using this technology in developing new drugs, aiming to provide essential theoretical insights and references for the application of related technologies in the field of new drug development.

Copyright of Practical Pharmacy & Clinical Remedies is the property of Editorial Department of Practical Pharmacy & Clinical Remedies and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

The effects of applying different types of organic fertilizer on neutral aroma component contents of upper and central leaves were studied with Zhongyan100 as the sample using randomized blocks design，and the data by using variance analysis，significance test and other methods were analyzed. The result showed that T2 resulted in the highest content of neophytadiene(1025.76 μ/g) in upper leaves，while T5 resulted in the highest content of neophytadiene (1322.02 μ/g) in middle leaves and also resulted in the greatest total carotenoid catabolites in all positions，the content of aromatic AA catabolites and cembratriendiol catabolites in two positions was the highest，and the greatest (24.76 μ/g) the total of mail lard reaction in upper leaves. Sensory smoking results indicated that T5 was the best in all of the treatments. After taste of their pure aroma，adequate aroma and comfortable，the T5 was more effective than other treatments to increase the content and harmony of aroma components. [ABSTRACT FROM AUTHOR]

Copyright of West China Journal of Stomatology is the property of Sichuan University, West China College of Stomatology and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

For effectively control of Mythimna separate Walker by reasonable application of emamectin benzoate, the cross-resistance of emamectin benzoate resistant population to other five insecticides and its biochemical resistance mechanisms were studied by bioassay and biochemical assay. The results revealed that medium level of cross-resistance to abamectin (resistance ratio:RR50=21.80), chlopyrifos (RR50=17.68) and methomyl (RR50=10.85), and low level of cross-resistance to phoxim (RR50=6.00) and cyfluthrin (RR50=5.65) for emamectin benzoate resistant population. The enzyme inhibitors piperonyl butoxide (PBO), diethyl maleate (DEM), S,S,S-tributyl phosphorotrithioate (DEF) and triphenyl phosphate (TPP) had significant synergism to emamectin benzoate in both resistant and susceptible populations. The contents of cytochrome P450 and b5, and activities of O-demethylase, glutathione S-transferases and carboxylesterase of the emamectin benzoate resistant population were 3.23, 3.65, 3.63, 1.64 and 2.66-fold of that in the susceptible population, respectively. These results indicated that the increased detoxification enzyme activities may play an important role in the resistance of M. separate to emamectin benzoate. [ABSTRACT FROM AUTHOR]

Protein kinase C (PKC) is a group of phospholipid-dependent serine/threonine protein kinases, which together with protein kinase A (PKA) and protein kinase G (PKG) constitute a superfamily of serine/threonine protein kinase AGC. PKC includes classic PKC, novel PKC, atypical PKC and some members of kinase associated with PKC (PRK). PKC is widely distributed in mammalian tissues and cells, which plays an important biological role in growth and metabolism, proliferation and differentiation of cells. A series of studies have demonstrated that variations of multiple cells, occurrence and development of diseases are related to the abnormal expression of PKC. Therefore, designing and searching for efficient PKC inhibitors have very important implications for synthesis of many kinds of effective drugs and treatment of a variety of clinical diseases including cancer, cardiovascular, and hypertension, et al. In recent years, the research on PKC inhibitors has become the focus of domestic and foreign research. A large number of literatures have reported many kinds of effective PKC inhibitors, and analyzed their function site, mechanism, clinical trial data and so on. The discovery of these PKC inhibitors has important implications for structural analysis of PKC and the treatment of diseases. So in this paper, the efficient PKC inhibitors are summarized. [ABSTRACT FROM AUTHOR]

In recent years, the number of advanced non-small cell lung cancer (NSCLC) patients has gradually increased, and the treatment methods have also been significantly increased. However, there are no standard treatment plans at home and abroad for third-line and above patients who are refractory to targeted therapy epidermal growth factor receptor (EGFR)/anaplastic lymphoma kinase (ALK) or chemotherapy. The clinical treatment effect is also not satisfactory. Anlotinib is a novel TKI targeting the vascular endothelial growth factor receptor (VEGFR), fibroblast growth factor receptor (FGFR), platelet-derived growth factor receptor (PDGFR) and c-Kit. ALTER0303 trail, phase III study has demonstrated that Anlotinib significantly prolonged overall survival (OS) and progression-free survival (PFS) in advanced NSCLC patients as 3rd line treatment. Here we report a case of advanced lung adenocarcinoma harboring KRAS mutation treated with Anlotinib. [ABSTRACT FROM AUTHOR]

Objective To observe the inhibitory and pro - apoptotic effects of two poly ( ADP - ribose ) polymerase ( PARP - 1 ) inhibitors, AG-014699 and AZD2281, on human hepatoma HepG2 cells and preliminarily explore the mechanism by which AG -014699 induces HepG2 cell apoptosis, and to provide a new therapeutic target for hepatoma. Methods The effects of different concentrations of AG - 014699 and AZD2281 on HepG2 cell proliferation were determined by MTT assay. The cell apoptosis rate was measured by flow cytometry. The expression levels of caspase - 3 and caspase - 8 were measured by Western Blot. Inter - group comparison was made by t test. Results Both AG -014699 and AZD2281 suppressed HepG2 cell proliferation in a time - and dose - dependent manner. However, the sensitivity of HepG2 cells to the two PARP - 1 inhibitors was diferent. The half - maximal inhibitory concentrations of AG - 014699 and AZD2281 at 48 h determined by MTT assay were about 20 μmol/L and 400 μmol/L, respectively. Flow cytometry and Western blot were not used to evaluate the apoptosis of HepG2 cells exposed to AZD2281 to which these cells were not sensitive. HepG2 cell apoptosis could be induced by 10, 30, and 50 μmol/L AG - 014699, and the highest apoptosis rate at 48h was significantly higher than that of the control group (31.00% ±2. 13% vs 0.9% ± 0.013%, P<0.01). Compared with those in the control group, the protein levels of caspase - 3 and caspase - 8 in HepG2 cells after 48 - h exposure to 30, and 50 μmol/L AG - 014699 increased. Conclusion The two PARP - 1 inhibitors AG-014699 and AZD2281 can inhibit the proliferation of HepG2 cells, which showed diferent sensitivities to the two inhibitors. AG - 014699 can induce HepG2 cell apoptosis by up - regulating the protein expression of caspase - 3 and caspase - 8. [ABSTRACT FROM AUTHOR]

The prevalence of osteoporosis has increased during recent years. The activation in the function of osteoclast is the main reason of osteoporosis. Cathepsin is expressed by osteoclast and involved in the degradation of collagen of hone, and causes osteoporosis. Cathepsin K is a kind of most important enzyme in the family of cathepsin. Odanacatib (ODN) is the inhibitor of cathepsin K, and it may be used to anti-osteoporosis thought inhibiting the degradation of bone collagen. It was found that the cortical thickness and bone minerals of cancellous increased after taking ODN in the studies, and then the density and the strength of bone increased. This study reviewed the pharmacological profile of ODN and the progresses of animal study and clinical trials about ODN. [ABSTRACT FROM AUTHOR]

To enhance rapeseed germplasm and improve rapeseed variety with resistance to tribenuron-methyl, seeds of Zhongshuang 9 (Brassica napus) were treated with 0.4%, 0.8%, 1.0% and 1.2% of EMS (ethyl methansulfonate) solution respectively. Results showed that (1)1. 0% of EMS was optional for mutagenesis and the phenotype variation in four M2 populations were 11.26%, 14.82%, 27.19% and 12.38% respectively. (2) Different concentrations of EMS had different effects on organs. (3) Three mutants with resistance to tribenuron -methyl were with mutation frequency about 0.0001. The M2 mutant library, including mutants of cotyledons, leaves, flowers, plant-types, siliques, was available for genetic studies and breeding in rapeseed. [ABSTRACT FROM AUTHOR]

To develop EP1 receptor antagonists with higher activities, the key factors that affect the activities of EPI antagonists were explored in this study. 103 EP1 antagonists were selected as data sets, and each molecule was calculated based on 254 parameters. Two regression methods of multiple linear regression (MLR) and principal component analysis (PCA) were used. The results show that the quantitative structure -activity relationship models using both the MLR and PCA exhibit good predictive ability. The statistical results by MLR show training set R² = 0.77, SEE = 0. 83, test set R² = 0.74, SEp = O. 33, and those by PCA show 2 training set R² = 0.72, SEE = 0. 45, test set R² = 0.71, SEP = 0.38. [ABSTRACT FROM AUTHOR]

The current standard treatment for hepatitis C virus (HCV) infection is a combination of pegylated interferon (PEG-IFN) and ribavirin. However, it remains incurable in some patients. In recent years, direct acting antiviral agents (DAAs) that target specific HCV enzymes in HCV life cycle have been developed rapidly. A combination of PEG-IFN, ribavirin, and DAAs can increase the sustained virological response rate in patients with chronic hepatitis C (CHC). For patients who cannot use or tolerate interferon therapy, a combination of various DAAs has good efficacy. Therefore, DAAs bring new hope for the treatment of CHC. [ABSTRACT FROM AUTHOR]

Melanin is an important factor affecting human skin color. This study defines its synthetic pathways and regulatory pathways have the practical significance for the treatment of diseases caused by pigmentation problems, such as chloasma. Besides, it also provides a theoretical basis for screening out melanin inhibitors and developing whitening and freckle products. At present, the melanin inhibitors used in whitening and freckle products mainly come from natural products and traditional Chinese medicine. This article first introduces the melanin biosynthesis pathway with tyrosinase as the core, defines the synthesis, transport and catalytic activity of tyrosinase as the three key factors affecting melanin synthesis, and then reviews two common types of melanin inhibitors, namely tyrosinase synthesis inhibitors and tyrosinase inhibitors derived from natural products and traditional Chinese medicine. This article provides guidance for the development of new melanin inhibitors, and puts forward the idea for combining and synergizing inhibitors according to different mechanisms, in order to develop new whitening formulas.

Objective: To investigate the effects of a novel polyamine metabolism enzyme inhibitor SI-4650 on autophagy and apoptosis of colon cancer CT-26 cells as well as their correlation., Methods: CT-26 cells treated with 40, 80 μmol·L -1 SI-4650 alone or in combination with 3-MA were used as experimental group. CT-26 cells treated with 0 μmol·L -1 SI-4650 alone or in combination with 3-MA were used as control group. Chemiluminescence was used to analyze the effect of SI-4650 on spermine oxidase (SMO) and acetylpolyamine oxidase(APAO) activity. High performance liquid chromatography (HPLC) was performed to detect cellular polyamine content.The CCK8 method was used to detect the inhibitory effect of SI-4650 on proliferation of CT-26 cells. PI single-staining/flow cytometry (FCM) were used to analyze cell cycle. Western blot were used to analyze autophagy. Apoptosis was analyzed by PI/FITC-Annexin V double staining, JC-1 fluorescent probe and Fluo-3 AM calcium ion fluorescent probe combined with flow cytometry and Western blot., Results: CCK8 assay showed that 24-,48-,72-hours treated with SI-4650 all could inhibit the proliferative activity of CT-26 cells in a dose- and time-dependent manner ( P ＜0.01) . The inhibition rate was 36.98% and 46.91% in 40 μmol·L -1 SI-4650 group and 80 μmol·L -1 SI-4650 group respectively. SI-4650 could significantly inhibit the activities of SMO and APAO interfere with polyamine metabolism and reduce the content of total polyamine in CT-26 cells ( P ＜0.01). SI-4650 could block CT-26 cells in G0/G1 phase, significantly reduce the number of cells in S phase( P ＜0.01), and lead to a significant increase in the contents of autophagy-related Beclin-1, LC3-II in CT-26 cells( P ＜0.01); At the same time, the concentration of calcium in CT-26 cells was increased, the mitochondrial membrane potential was decreased, the expressions of c-PARP and Bax were increased, the content of Bcl-2 was decreased, and the number of apoptotic cells was increased. After SI-4650 combined with autophagy inhibitor 3-MA treatment of CT-26 cells, the level of autophagy, the apoptosis-related protein, mitochondrial membrane potential and calcium ion concentration were decreased, and the number of apoptotic cells was decreased., Conclusion: SI-4650 has the pharmacological activity of killing colon cancer CT-26 cells, and its mechanism may be related to the interference of polyamine metabolism and induction of cell apoptosis and autophagy. In this process, autophagy is inhibited to block apoptosis, autophagy and apoptosis combined to kill tumor cells.

As aberrant enzyme functions are often closely associated with various diseases, enzymes represent one of the most important drug target groups. Historically, natural products have been the most productive sources for drug discovery. However, the complexity of the components leads to a bottleneck in separation and purification, thus restricting activity-based drug screening. Due to the unique advantages such as high separation efficiency and without sample matrix interference, the advantage of capillary electrophoresis (CE) directly applied to extracts without purification for activity-based drug screening is of great attractive. In this review, we have summarized the application of CE for screening enzyme inhibitors in natural products during the last decade. In particular, we discuss important examples for screening inhibitors of transferases (kinases), hydrolases, oxidoreductases, etc. We also present an overview of the technical details concerning the separation modes in CE and enzyme kinetics. Finally, we provide future prospects for the application of CE to the screening of active ingredients in natural products.

Continuing advances in dentin bonding technology and adhesives revolutionized bonding of resin-based composite restorations. However, hybrid layers created by contemporary dentin adhesives present imperfect durability, and degradation of collagen matrix by endogenous enzymes is a significant factor causing destruction of hybrid layers. Bond durability can be improved by using enzyme inhibitors to prevent collagen degradation and to preserve integrity of collagen matrix. This review summarizes progress on matrix metalloproteinase inhibitors (including chlorhexidine, ethylenediaminetetraacetic acid, quaternary ammonium salt, tetracycline and its derivatives, hydroxamic acid inhibitors, bisphosphonate derivative, and cross-linking agents) and suggests prospects for these compounds.

Objective: To analyze the susceptibility of influenza A (H3N2) viruses to neuraminidase inhibitors during 2011-2012 in Mainland China., Methods: All the tested viruses were obtained from the Chinese National Influenza Surveillance Network, which covers 31 provinces in mainland China, including 408 network laboratories and 554 sentinel hospitals. In total 1 903 viruses were selected with isolation date from January 1, 2011 to December 31, 2012 in Mainland China, among these viruses, 721 were confirmed to be influenza A (H3N2) virus by Chinese National Influenza Center and tested for the susceptibility to oseltamivir and zanamivir using chemiluminescence-based assay. The neuraminidase inhibitor sensitive reference virus A/Washington/01/2007 (119E) and oseltamivir resistant virus A/Texas/12/2007 (E119V) were used as control in this study. The t -test was used to compare the difference of NAI susceptibility of viruses isolated from different years., Results: The half maximal inhibitory concentration (IC₅₀) of A/Washington/01/2007 for oseltamivir and zanamivir was (0.10 ± 0.02) and (0.30 ± 0.05) nmol/L, respectively. The IC₅₀ of A/Texas/12/2007 for oseltamivir and zanamivir was (4.27 ± 1.60) and (0.20 ± 0.03) nmol/L, respectively. Among the 721 influenza A (H3N2) viruses, 132 influenza A (H3N2) viruses were isolated in 2011 and 589 influenza A (H3N2) viruses were isolated in 2012. The IC50 for oseltamivir ranged from 0.04 to 0.62 nmol/L for viruses isolated in 2011 and ranged from 0.02 to 0.95 nmol/L for viruses in 2012, and the IC₅₀ of all the viruses tested was within 10-fold IC₅₀ (1.0 nmol/L) of the neuraminidase inhibitor sensitive reference virus A/Washington/01/2007. The IC50 of zanamivir ranged from 0.12 to 0.80 nmol/L for viruses in 2011 and ranged from 0.04 to 0.72 nmol/L for viruses in 2012, and was within 10-fold IC₅₀ (3.0 nmol/L) of the neuraminidase inhibitor sensitive reference virus A/Washington/01/2007., Conclusion: The influenza A(H3N2) viruses isolated during 2011-2012 in Mainland China were tested to be sensitive to oseltamivir and zanamivir.

Objective: To investigate the effect of selective phosphatase inhibitors Salubrinal on autophagy and apoptosis in the lung tissue of rats with acute paraquat (PQ) poisoning, and to explore its mechanism., Methods: 200 Wistar rats were randomly divided into four groups by randomized arrangement table formed by computer, with 50 rats in each group. PQ poisoning model was reproduced by one time gastric lavage with 1 mL of 40 mg/kg PQ solution followed by intraperitoneal injection of 1 mL normal saline (NS) once a day. The rats in control group were lavaged once with 1 mL of NS followed by intraperitoneal injection of 1 mL NS twice a day. The rats in Sal 0.5 and Sal 1.0 groups were intraperitoneal injected with 1 mL Salubrinal 0.5 mg/kg or 1.0 mg/kg on the 1st, 3rd, and 5th day after PQ poisoning once a day. The lung tissue was harvested on the 7th day after poisoning, and the changes in histomorphology were observed using hematoxylin and eosin (HE) staining. The positive expression of autophagy-related protein LC3-II in lung tissue was observed after immunohistochemistry staining, and LC3-II and caspase-3 protein expressions were determined by Western Blot., Results: HE staining results showed partial abnormal pulmonary structure in the PQ poisoning group: collapse of pulmonary alveoli, enlargement of the cavity, local infiltration of inflammatory cells, increasing thickness in the alveoli wall and obvious bleeding in the local lung tissue. Compared with the PQ poisoning group, the above changes in Sal 0.5 and Sal 1.0 groups were obviously relieved. It was shown by immunohistochemistry staining that compared with control group, the positive expression of LC3-II was obviously decreased in the PQ poisoning group, Sal 0.5, and Sal 1.0 groups (A value: 78.34 ± 10.71, 76.52 ± 8.21, 77.48 ± 9.11 vs. 117.58 ± 15.26, all P<0.05). There was no significant difference in positive expression of LC3-II between each of the later three groups (all P>0.05). Western Blot results showed: compared with the control group, the protein expressions of LC3-II and caspase-3 were significantly increased in PQ poisoning group [LC3-II (A value): 0.22 ± 0.05 vs. 0.14 ± 0.03, caspase-3 (A value): 0.115 ± 0.013 vs. 0.023 ± 0.006, both P<0.05]. Compared with PQ poisoning group, the protein expressions of LC3-II and caspase-3 were obviously decreased in the Sal 0.5 and Sal 1.0 groups [LC3-II (A value): 0.19 ± 0.05, 0.18 ± 0.04 vs. 0.22 ± 0.05; caspase-3(A value): 0.078 ± 0.012, 0.076 ± 0.010 vs. 0.115 ± 0.013, all P<0.05]., Conclusions: The endoplasmic reticulum stress-autophagy is activated in the pulmonary cell of acute PQ poisoning rats. Salubrinal can decrease the autophagy and apoptosis in the lung of rats with acute PQ poisoning, which play a role in the treatment.

Objective: To evaluate the effect of combined targeting of MEK and PI3K signaling pathways on K-ras mutated non-small cell lung cancer cell line A549 cells and the relevant mechanisms., Methods: A549 cells were treated with different concentrations of two inhibitors. Growth inhibition was determined by MTT assay. According to the results of MTT test, the cells were divided into four groups: the control group, PI3K inhibitor group (GDC-0941,0.5 and 5.0 µmol/L), combination group I (0.5 µmol/L AZD6244+0.5 µmol/L GDC-0941) and combination group II (5.0 µmol/L AZD6244+5.0 µmol/L GDC-0941). The cell cycle and apoptosis were analyzed by flow cytometry. The expression of proteins related to apoptosis was tested with Western blot., Results: Both GDC-0941 and AZD6244 inhibited the cell proliferation. The combination group II led to a stronger growth inhibition. The combination group I showed an antagonistic effect and combination group II showed an additive or synergistic effect. Compared with the control group, the combination group I led to reduced apoptotic rate [(20.70 ± 0.99)% vs. (18.65 ± 0.92 )%, P > 0.05]; Combination group II exhibited enhanced apoptotic rate [(37.85 ± 3.18)% vs. (52.27 ± 4.36)%, P < 0.01]. In addition, in the combination group II, more A549 cells were arrested in G0/G1 phase and decreased S phase (P < 0.01), due to the reduced expressions of CyclinD1 and Cyclin B1, the increased cleaved PARP and the diminished ratio of Bcl-2/Bax., Conclusions: For single K-ras mutated NSCLC cell line A549 cells, combination of RAS/MEK/ERK and PI3K/AKT/mTOR inhibition showed synergistic effects depending on the drug doses. Double pathways targeted therapy may be beneficial for these patients.

Objective: To investigate the effect of Leptin on differentiation of nueral stem cell and explore the underline molecular mechanism., Methods: Co-culture neural stem cell with different concentration of Leptin (0, 0.05, 0.2, 0.5, 1, 2 mg/L), the effect of Leptin on differentiation of nueral stem cell was analyzed. After treated the nueral stem cell with inhibitors of Jak-STAT3 (Janus family tyrosine kinases (Jak kinases) and signal transducers and activators of transcription (STAT proteins) and PI3k-Akt (phosphatidylinositol-3-kinase (PI3K)/protein kinase B (Akt)), the STAT3, phosphorylated-STAT3, Akt, phosphorylated-Akt levels and GFAP were detected with western blot and fluoroimmunoassay respectively., Results: Morphology observation found nueral stem cell to change to astrocyte. An increased astrocyte marker (GFAP) in Leptin group but not Tuj-1 or MBP with fluoroimmunoassay and western blot detection was observed. The expression of GFAP began at 24 h after co-culture, and increased consistent with the concentration of Leptin. Jak-STAT3 inhibitors but not PI3k-Akt inhibitors decreased the expression of phosphorylated-STAT3, and accompanily decreased the expression of GFAP., Conclusion: Leptin may have an effect on the astroglial differentiation on neural stem cells through the JAK-STAT3 pathway but not PI3k-Akt pathway.

Combined with molecular docking model, a fluorescence method was applied to investigate the interaction between quercetin and beta-glucosidase and the acting mechanism. The interaction between beta-glucosidase and quercetin, as well as the enzyme inhibitor 4-nitrophenyl-beta-D-thioglucoside, was studied by the AutoDock4.2 molecular docking model, respectively. The binding reaction was simultaneously studied using fluorescence quenching method. The results showed that these interactions result in the endogenous fluorescence quenching of beta-glucosidase, which belongs to a static quenching mechanism. The calculated binding constants were 4.36 X 10(4), 4.04 x 10(4) and 3.18 x 10(4) L mol(-1) at 17, 27 and 37 degrees C, respectively. The results revealed that quercetin tended to bind with beta-glucosidase mainly by hydrogen bond and hydrophobic interaction, as well as electrostatic forces. Both fluorescence spectroscopy and molecular docking are complementary to each other for the investigation of the interaction between beta-glucosidase and quercetin from the experimental and theoretical view.

Objective: To determine the effects of Tongxinluo on cell viability and tissue factor (TF) in AngII induced vascular endothelial cells and to investigate its mechanism., Methods: AngII(10(-6)mol/L) was added to human vascular endothelial cells (HUVECs) culture media alone or with various concentration of Tongxinluo drug containing plasma (5%,10%, and 20%) added 30 minutes before AngII. Cell viability was evaluated after 24-hour incubation with AngII in a dose manner. TF, AngII type 1 receptor (AT(1)) mRNA, NO synthase (NOS) and NO were observed after 24-hour incubation with AngII. In addition, NOS inhibitor nomega-nitro-larginine (L-NAME) was added 30 minutes before Tongxinluo and AngII. Cell viability, TF, AT(1)mRNA, the level of NOS and NO were evaluated after 24-hour incubation with Tongxinluo and AngII., Results: Tongxinluo significantly improved AngII induced endothelial cell viability and the effect was the most obvious at 10%. Tongxinluo (10%) decreased the TF and AT(1) mRNA while increased the NOS and NO levels. L-NAME obviously inhibited the effects of Tongxinluo on cell viability, TF, AT(1) mRNA, and NOS and NO levels., Conclusion: Up-regulating NOS-NO signaling may be the mechanism of Tongxinluo on cell viability and TF in AngII induced vacular endothelial cells.

Objective: To build 3D-pharmacophore model of epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors using distance comparisons method and design novel EGFR inhibitors., Methods: Thirteen typical EGFR inhibitors were selected, and their biologically active conformations were obtained by using DOCK5.0 program, then 3D-pharmacophore model of EGFR inhibitors was built using distance comparisons method., Results: Validation of the 3D-pharmacophore model was carried out and novel structures with potential inhibitory activity were selected by means of 3D-searching and docking method., Conclusion: This method can improve hit rate of lead compounds discovery and can be used to design novel EGFR inhibitors.

Objective: Nitric oxide (NO) precursor L-arginine (L-Arg) and NOS inhibitor NG-nitro-L-arginine methyl ester (L-NAME) were used to investigate the role of endogenous NO in airway hyperresponsiveness of asthmatic rats., Methods: Asthmatic wistar rats were developed by sensitization and challenging with ovalbumin. Airway responsivess to acetylcholine (Ach) was measured in vitro in isolated and perfused rat tracheal rings with or without epithelium after inhibiting or stimulating NO synthesis., Results: After asthmatic rat tracheal rings were incubated in vitro with L-NAME 10(-5) mol/L, the maximal response of tracheal rings to Ach was increased compared with that of the control group. When asthmatic rat tracheal rings were coincubated with L-Arginine 2 x 10(-5) mol/L and L-NAME 10(-5) mol/L, the maximal response was decreased compared with that of the L-NAME group. Incubation with higher concentrations of L-Arginine did not change asthmatic rat tracheal responsiveness to Ach. Epithelium-denuded tracheal rings in the asthmatic rats showed a significant (P < 0.05) upward shift in the Ach concentration response curve; the maximal response was increased compared with the tracheal intact group. In the denuded tracheal rings, L-NAME and L-Arginine did not influence the shape of the Ach concentration-response curve and the maximal response., Conclusions: Endogenous nitric oxide, a mediator of bronchodilation, is synthesized from L-Arginine by NO synthase in asthmatic rats. The epithelium lining airway smooth muscle probably contributes to nitric oxide synthesis. Our finding suggests that the airway in asthmatic rats could not use exogenous L-Arginine to synthesize nitric oxide.