TFAP2A upregulates SKA3 to promote glycolysis and reduce the sensitivity of lung adenocarcinoma cells to cisplatin.

INTRODUCTION: This work was designed to delve into the effects of SKA3 on glycolysis and cisplatin (CDDP) resistance in LUAD cells and to find new possibilities for individualized treatment of LUAD. METHODS: LUAD mRNA expression data from the TCGA database were procured to scrutinize the differential expression patterns of SKA3 in both tumor and normal tissues. GSEA and Pearson correlation analyses were employed to elucidate the impact of SKA3 on signaling pathways within the context of LUAD. In order to discern the upstream regulatory mechanisms, the ChEA and JASPAR databases were utilized to predict the transcription factors and binding sites associated with SKA3. qRT-PCR and Western blot were implemented to assay the mRNA and protein expression levels of SKA3 and TFAP2A. Chromatin immunoprecipitation (ChIP) and dual luciferase assays were performed to solidify the binding relationship between the two. Extracellular acidification rate, glucose consumption, lactate production, and glycolysis-related proteins (HK2, GLUT1, and LDHA) were used to evaluate the level of glycolysis. Cell viability under CDDP treatment was determined utilizing the CCK-8, allowing for the calculation of IC50. The expression levels of SKA3 and TFAP2A proteins were detected by immunohistochemistry (IHC). RESULTS: SKA3 exhibited upregulation in LUAD tissues and cell lines, establishing a direct linkage with glycolysis pathway. Overexpression of SKA3 fostered glycolysis in LUAD, resulting in reduced sensitivity towards CDDP treatment. The upstream transcription factor of SKA3, TFAP2A, was also upregulated in LUAD and could promote SKA3 transcription. Overexpression of TFAP2A also fostered the glycolysis of LUAD. Rescue assays showed that TFAP2A promoted glycolysis in LUAD cells by activating SKA3, reducing the sensitivity of LUAD cells to CDDP. The IHC analysis revealed a positive correlation between high expression of SKA3 and TFAP2A and CDDP resistance. CONCLUSION: In summary, TFAP2A can transcriptionally activate SKA3, promote glycolysis in LUAD, and protect LUAD cells from CDDP treatment, indicating that targeting the TFAP2A/SKA3 axis may become a plausible and pragmatic therapeutic strategy for the clinical governance of LUAD.

VHL suppresses autophagy and tumor growth through PHD1-dependent Beclin1 hydroxylation.

The Von Hippel-Lindau (VHL) protein, which is frequently mutated in clear-cell renal cell carcinoma (ccRCC), is a master regulator of hypoxia-inducible factor (HIF) that is involved in oxidative stresses. However, whether VHL possesses HIF-independent tumor-suppressing activity remains largely unclear. Here, we demonstrate that VHL suppresses nutrient stress-induced autophagy, and its deficiency in sporadic ccRCC specimens is linked to substantially elevated levels of autophagy and correlates with poorer patient prognosis. Mechanistically, VHL directly binds to the autophagy regulator Beclin1, after its PHD1-mediated hydroxylation on Pro54. This binding inhibits the association of Beclin1-VPS34 complexes with ATG14L, thereby inhibiting autophagy initiation in response to nutrient deficiency. Expression of non-hydroxylatable Beclin1 P54A abrogates VHL-mediated autophagy inhibition and significantly reduces the tumor-suppressing effect of VHL. In addition, Beclin1 P54-OH levels are inversely correlated with autophagy levels in wild-type VHL-expressing human ccRCC specimens, and with poor patient prognosis. Furthermore, combined treatment of VHL-deficient mouse tumors with autophagy inhibitors and HIF2α inhibitors suppresses tumor growth. These findings reveal an unexpected mechanism by which VHL suppresses tumor growth, and suggest a potential treatment for ccRCC through combined inhibition of both autophagy and HIF2α.

ALDOB/KAT2A interactions epigenetically modulate TGF-ß expression and T cell functions in hepatocellular carcinogenesis.

BACKGROUND AND AIMS: Cross talk between tumor cells and immune cells enables tumor cells to escape immune surveillance and dictate responses to immunotherapy. Previous studies have identified that downregulation of the glycolytic enzyme fructose-1,6-bisphosphate aldolase B (ALDOB) in tumor cells orchestrated metabolic programming to favor HCC. However, it remains elusive whether and how ALDOB expression in tumor cells affects the tumor microenvironment in HCC. APPROACH AND RESULTS: We found that ALDOB downregulation was negatively correlated with CD8 + T cell infiltration in human HCC tumor tissues but in a state of exhaustion. Similar observations were made in mice with liver-specific ALDOB knockout or in subcutaneous tumor models with ALDOB knockdown. Moreover, ALDOB deficiency in tumor cells upregulates TGF-ß expression, thereby increasing the number of Treg cells and impairing the activity of CD8 + T cells. Consistently, a combination of low ALDOB and high TGF-ß expression exhibited the worst overall survival for patients with HCC. More importantly, the simultaneous blocking of TGF-ß and programmed cell death (PD) 1 with antibodies additively inhibited tumorigenesis induced by ALDOB deficiency in mice. Further mechanistic experiments demonstrated that ALDOB enters the nucleus and interacts with lysine acetyltransferase 2A, leading to inhibition of H3K9 acetylation and thereby suppressing TGFB1 transcription. Consistently, inhibition of lysine acetyltransferase 2A activity by small molecule inhibitors suppressed TGF-ß and HCC. CONCLUSIONS: Our study has revealed a novel mechanism by which a metabolic enzyme in tumor cells epigenetically modulates TGF-ß signaling, thereby enabling cancer cells to evade immune surveillance and affect their response to immunotherapy.

Electrocatalytic hydrogen evolution with a copper porphyrin bearing meso-(o-carborane) substituents.

A newly designed copper complex of 5,15-bis(pentafluorophenyl)-10,20-bis(o-carborane)porphyrin (1) was synthesized and tested for the electrocatalytic hydrogen evolution reaction (HER). In acetonitrile, 1 was much more efficient than Cu 5,15-bis(pentafluorophenyl)-10,20-diphenylporphyrin (2) for electrocatalytic HER by shifting the catalytic wave to the anodic direction by 190 mV. In aqueous media, 1 also outperformed 2 by achieving higher current densities under smaller overpotentials. This enhancement was attributed to the aromatic and the strong electron-withdrawing properties of o-carborane groups. This work is significant to address the crucial effects of meso-(o-carborane) substituents of metal porphyrins on boosting the electrocatalytic HER.

[Research progress on the effect mechanism of acupuncture-moxibustion for pressure injuries].

Evidence shows that acupuncture-moxibustion could promote the healing of pressure injuries (PI), but its action mechanism is not fully understood. This review summarizes the basic research literature of acupuncture-moxibustion for PI and identifies that the mechanism of acupuncture-moxibustion for PI is related with regulation of related signaling pathway target proteins, improvement of inflammatory response, modulation of vascular microenvironment, attenuation of oxidative stress damage, and inhibition of cell apoptosis. The review also points out the current limitations and future research directions. It emphasizes the need for further exploration of the upstream regulatory mechanism, specific cellular molecules, and the interactions among these molecules. A multi-level, multi-target, and multi-dimensional approach is required to fully understand the mechanism underlying the promotion of PI healing by acupuncture-moxibustion.

Endoscopy-guided intraoral extraction of ectopic mandibular third molar in the subcondylar region planned with three-dimensional imaging: A case report and literature review.

Ectopic mandibular third molar (EMTM) in the subcondylar region is a rare clinical condition, especially for a subtype confined between the mandibular foramen and condylar neck. The etiology is currently uncertain and the optimal management of this specific subtype remains not well defined. We reported a case of this specific subtype of EMTM that was minimally invasively extracted by endoscopy-guided intraoral surgery, planned preoperatively using three-dimensional (3D) imaging of cone beam computed tomography (CBCT), with no complications postoperatively caused by the routine surgery. We also reviewed nine relevant literature to expand the clinical features and therapeutic management of this specific subtype of EMTM. Etiologically, persistent cystic pressure may be a major cause of EMTM displaced into the subcondylar region. For extraction of this specific EMTM, the combination of 3D CBCT-based imaging and endoscopy-assisted intraoral minimally invasive surgery could be considered as the priority option for patients without facial fistula.

Interfacially Polymerized ß-Cyclodextrin Film for Rapid Removal of Organic Dyes.

Porous cyclodextrin-based polymers are widely used for the rapid removal of organic pollutants in water. Traditional cyclodextrin-based polymers are prepared in the organic phase, which is time consuming and costly. Herein, a novel cyanuricchloride (TCT) cross-linked porous ß-cyclodextrin-based thin-film composite membrane is designed in the aqueous phase by interfacial polymerization. A self-standing TCT-CDP film is formed instantly at the surface of water phase at room temperature. Several different water-soluble organic dyes such as Methylene Blue, Neutral Red, Auramine, Brilliant Green, and Crystal Violet are selected for rejection study with TCT-CDP membrane. The effective rejection of TCT-CDP membrane for typical dyes is up to 99%, indicating TCT-CDP membrane exhibit excellent selectivity for separation of organic dyes from water.

Metabolomic and transcriptomic analyses provide insights into variations in flavonoids contents between two Artemisia cultivars.

BACKGROUND: Plants in the genus Artemisia are rich in active ingredients and specialized metabolites. Many of these compounds, especially flavonoids, have potential medicinal and nutritional applications, and are of growing interest to scientists due to their wide range of pharmacological and biological activities. Artemisia cultivars are commonly used as raw materials for medicine, food, and moxibustion in China. However, most of the metabolites produced by Artemisia species have not been identified, and few studies have addressed differences in active compounds between species and cultivars. RESULTS: We here investigated two Artemisia cultivars, 'Nanyangshiyong' (NYSY) and 'Nanyangyaoyong' (NYYY), which are commonly used in foods and moxibustion, respectively. NYSY and NYYY were confirmed to be Artemisia argyi cultivars. Total flavonoids contents and antioxidant activities were higher in NYYY than in NYSY. A total of 882 metabolites were identified in the samples; most of the potentially medicinally active compounds, especially flavonoids (e.g., flavone, flavonol, isoflavone, and anthocyanin), were up-regulated in NYYY compared to NYSY. Furthermore, most of the genes related to flavonoids biosynthesis were up-regulated in NYYY. Correlation analysis was used to identify putative members of transcription factor families that may regulate genes encoding key flavonoids biosynthesis enzymes. CONCLUSIONS: We found that the antioxidant activities and flavonoids contents significantly varied between two Artemisia cultivars of the same species. We also uncovered metabolomic and transcriptomic evidence of the molecular phenomena underlying those differences in flavonoids contents between the two Artemisia cultivars. This study provides a wealth of data for future utilization and improvements of Artemisia cultivars, and highlights a need to study the specific metabolite profiles of plants that are used in foods and medicines.

Prognosis of adenoid cystic carcinoma in head and neck region treated with different regimens-A single-centre study.

BACKGROUND: No study has evaluated the impact of regimen on recurrence, metastasis and survival in patients with adenoid cystic carcinoma (ACC). The present study aimed to compare the efficacy of radioactive seed implantation and other regimens in treating ACC, so as to investigate the clinical applicability of radioactive seed implantation and determine the indications for this regimen. METHODS: A total of 188 patients with ACC in oromaxillofacial region were allocated to four groups according to the treatment regimen: group 1 was treated with a combination of surgery and 125 I seed therapy, group 2 with a combination of surgery and external radiotherapy, group 3 with surgery, whereas group 4 was untreated. The Kaplan-Meier method was used to assess the survival rates, and the Cox regression analyses were used to identify the associated prognostic factors. RESULTS: The overall survival rates of 188 patients and groups 1, 2, 3 and 4 were 85.7%, 75%, 68.2% and 37.5%, respectively. Cox regression analysis revealed that age, T stage, N stage and regimen were independent prognostic factors of survival. Amongst patients with primary ACC, the efficacy of radioactive seed implantation was higher in those with perineural invasion than in those without. CONCLUSION: Patient age, T stage, N stage and regimen are independent prognostic factors of survival in patients with ACC. Patients treated with surgery combined with postoperative 125 I seed radiotherapy have a higher overall survival rate, and those with perineural invasion are more suitable for radioactive seed implantation therapy.

Quantitative fingerprint and antioxidative properties of Artemisia argyi leaves combined with chemometrics.

Quantitative fingerprint and differences of Artemisia argyi from different varieties, picking time, aging year, and origins were analyzed combing with chemometrics. The antioxidant activity was determined and antioxidant markers of Artemisia argyi were screened. Variety WA3 was significantly different from that of the other varieties. Fingerprint peak response and antioxidant activity of A. argyi picked in December were lower than samples collected in May and August. Fresh A. argyi leaves were significantly superior to withered leaves and stems. Artemisia argyi aging 1-5 years presented a classification trend. Antioxidant activity of A. argyi produced in Nanyang was generally superior to others origins. Peak 9, isochlorogenic acid A, and 6-methoxyluteolin contributed greatly for classification of A. argyi from different variety, picking time, aging year, and origin. Isochlorogenic acid A, isochlorogenic acid C, 6-methoxyluteolin, and chlorogenic acid were selected as antioxidant marker of A. argyi. The method based on quantitative fingerprint, antioxidant activity evaluation, and chemometrics was reliable to analyze the differences of A. argyi samples from different sources.

SRGAP2 controls colorectal cancer chemosensitivity via regulation of mitochondrial complex I activity.

Mitochondrial respiration and metabolism play an important role in the occurrence and development of colorectal cancer (CRC). In this study, we identified a functional pool of SLIT-ROBO Rho GTPase-activating protein 2 (SRGAP2) in the mitochondria of CRC cells as an important regulator of CRC chemosensitivity. We found that SRGAP2 levels were increased in CRC cells in comparison to normal colorectal cells. Loss of mitochondrial SRGAP2 led to significant decrease in mitochondrial respiration and strongly sensitized the CRC cells to chemotherapy drugs. Mechanistically, SRGAP2 physically interacts with mitochondrial complex I and positively modulates its activity. In particular, chemosensitization upon SRGAP2 loss was phenocopied by the treatment of complex I inhibitor. Thus, our results demonstrate that SRGAP2 functions as a key regulator of CRC chemosensitivity, identifying SRGAP2 as a promising therapeutic target to enhance the efficacy of chemotherapy in CRC.

Myostain is involved in ginsenoside Rb1-mediated anti-obesity.

CONTEXT: Obesity, one of the major public health problems worldwide, has attracted increasing attention. Ginsenoside Rb1 is the most abundant active component of Panax ginseng C.A.Mey (Araliaceae) and is reported to have beneficial effects on obesity and diabetes. However, the mechanisms by which Rb1 regulates obesity remain to be explored. OBJECTIVE: This paper intends to further explore the mechanism of Rb1 in regulating obesity. MATERIALS AND METHODS: The C57BL/6 obese mice were divided into two groups: the control (CTR) and Rb1. The CTR group [intraperitoneally (ip) administered with saline] and the Rb1 group (ip administered with Rb1, 40 mg/kg/d) were treated daily for four weeks. In vitro, Rb1 (0, 10, 20, 40 µM) was added to differentiated C2C12 cells and Rb1 (0, 20, 40 µM) was added to 3T3-L1 cells. After 24 h, total RNA and protein from C2C12 cells and 3T3-L1 cells were used to detect myostatin (MSTN) and fibronectin type III domain-containing 5 (FNDC5) expression. RESULTS: Rb1 reduced the body weight and adipocyte size. Improved glucose tolerance and increased basic metabolic activity were also found in Rb1 treated mice. MSTN was downregulated in differentiated C2C12 cells, 3T3-L1 cells and adipose tissues upon Rb1 treatment. FNDC5 was increased after Rb1 treatment. However, MSTN overexpression attenuated Rb1-mediated decrease accumulation of lipid droplets in differentiated 3T3-L1 adipocytes. DISCUSSION & CONCLUSIONS: Rb1 may ameliorate obesity in part through the MSTN/FNDC5 signalling pathway. Our results showed that Rb1 can be used as an effective drug in the treatment of human obesity.

Non-Coding RNA Analyses of Seasonal Cambium Activity in Populus tomentosa.

Non-coding RNA, known as long non-coding RNA (lncRNA), circular RNA (circRNA) and microRNA (miRNA), are taking part in the multiple developmental processes in plants. However, the roles of which played during the cambium activity periodicity of woody plants remain poorly understood. Here, lncRNA/circRNA-miRNA-mRNA regulatory networks of the cambium activity periodicity in Populus tomentosa was constructed, combined with morphologic observation and transcriptome profiling. Light microscopy and Periodic Acid Schiff (PAS) staining revealed that cell walls were much thicker and number of cell layers was increased during the active-dormant stage, accompanied by abundant change of polysaccharides. The novel lncRNAs and circRNAs were investigated, and we found that 2037 lncRNAs and 299 circRNAs were differentially expression during the vascular cambium period, respectively. Moreover, 1046 genes were identified as a target gene of 2037 novel lncRNAs, and 89 of which were the miRNA precursors or targets. By aligning miRNA precursors to the 7655 lncRNAs, 21 lncRNAs were identified as precursors tof 19 known miRNAs. Furthermore, the target mRNA of lncRNA/circRNA-miRNA network mainly participated in phytohormone, cell wall alteration and chlorophyll metabolism were analyzed by GO enrichment and KEGG pathway. Especially, circRNA33 and circRNA190 taking part in the phytohormone signal pathway were down-regulated during the active-dormant transition. Xyloglucan endotransglucosylase/hydrolase protein 24-like and UDP-glycosyltransferase 85A1 involved in the cell wall modification were the targets of lncRNA MSTRG.11198.1 and MSTRG.1050.1. Notably, circRNA103 and MSTRG.10851.1 regulate the cambium periodicity may interact with the miR482. These results give a new light into activity-dormancy regulation, associated with transcriptional dynamics and non-coding RNA networks of potential targets identification.

The mechanism of oxymatrine on atopic dermatitis in mice based on SOCS1/JAK-STAT3 pathway.

Based on the suppressor of cytokine signaling 1 (SOCS1)/Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) pathway, the mechanism of oxymatrine in the treatment of atopic dermatitis (AD) was preliminarily explored in this study. C57BL/6 mice were induced to establish AD model by smearing carbotriol (MC903) on their back. The AD mice were randomly divided into model group, oxymatrine groups with three dosages (25, 50 and 100 mg/kg), (n = 10). Oxymatrine groups were intragastric administered once daily for 14 days. The same volume of saline was given in the normal control group and model group once daily for 14 days. Subsequently, HE staining was used to observe the pathological changes of skin tissue, ELISA was used to detect the levels of serum inflammatory factors including interleukin-4, 6 and 17 (IL-4, IL-6, and IL-17), tumor necrosis factor-α (TNF-α) and immunoglobulin E (IgE). Immunohistochemistry was used to detect the expression of suppressor of cytokine signaling 1 and CD3 in skin tissue, and Western blotting was used to detect the proteins in suppressor of cytokine signaling 1/JAK-STAT3 pathway. Compared with the normal control group, the pathological damage of mice in the model group, such as skin hyperplasia, edema, congestion and inflammatory infiltration, aggravated increased significantly. And the expression of serum inflammatory factors, CD3 positive expression and JAK-STAT3 pathway protein in the model group were increased (p < .05), and the expression of suppressor of cytokine signaling 1 protein (p < .05) was decreased. Compared with the model group, the above pathological damage of the mice was reduced, and the serum inflammatory factors, JAK-STAT3 pathway protein, and CD3 positive expression were decreased as a dose-dependant manner (p < .05), and the expression of suppressor of cytokine signaling 1 protein was increased as a dose-dependent manner (p < .05). Oxymatrine can improve the skin inflammation symptoms of AD mice by up regulating the expression of suppressor of cytokine signaling 1, inhibiting the activation of JAK-STAT3 pathway and blocking the activation of T lymphocytes.

Degradation of 2,4-dichlorophenol by cathodic microarc plasma electrolysis: characteristics and mechanisms.

In this work, we used cathodic microarc plasma electrolysis (CMPE) to degrade 2,4-dichlorophenol (2,4-DCP) in simulated wastewater. By investigating and comparing the removal efficiencies and chemical oxygen demand (COD) during the degradation process, higher bath voltage and alkaline condition were considered as more suitable for the 2,4-DCP decomposition. Higher initial 2,4-DCP concentration was attributed to the increase in the utilisation of the energy input. The plasma characteristics during CMPE were studied by optical emission spectroscopy (OES). It was found that the 2,4-DCP directly participated in the plasma discharge process. Furthermore, by studying the evolution of intermediate products at different experimental parameters, it was found that the existence of Cl- played an important role in the opening of benzene ring, which activated the ortho-substitutions of hydroxyl, meanwhile accelerated the p-substitutions. The instantaneous high temperature and high pressure and the Cl- that were generated and driven by cathodic plasma made the decomposition of 2,4-DCP much quicker.

PTK7-Targeting CAR T-Cells for the Treatment of Lung Cancer and Other Malignancies.

In spite of impressive success in treating hematologic malignancies, adoptive therapy with chimeric antigen receptor modified T cells (CAR T) has not yet been effective in solid tumors, where identification of suitable tumor-specific antigens remains a major obstacle for CAR T-cell therapy due to the "on target off tumor" toxicity. Protein tyrosine kinase 7 (PTK7) is a member of the Wnt-related pseudokinases and identified as a highly expressed antigen enriched in cancer stem cells (CSCs) from multiple solid tumors, including but not limited to triple-negative breast cancer, non-small-cell lung cancer, and ovarian cancer, suggesting it may serve as a promising tumor-specific target for CAR T-cell therapy. In this study, we constructed three different PTK7-specific CAR (PTK7-CAR1/2/3), each comprising a humanized PTK7-specific single-chain variable fragment (scFv), hinge and transmembrane (TM) regions of the human CD8α molecule, 4-1BB intracellular co-stimulatory domain (BB-ICD), and CD3ζ intracellular domain (CD3ζ-ICD) sequence, and then prepared the CAR T cells by lentivirus-mediated transduction of human activated T cells accordingly, and we sequentially evaluated their antigen-specific recognition and killing activity in vitro and in vivo. T cells transduced with all three PTK7-CAR candidates exhibited antigen-specific cytokine production and potent cytotoxicity against naturally expressing PTK7-positive tumor cells of multiple cancer types without mediating cytotoxicity of a panel of normal primary human cells; meanwhile, in vitro recursive cytotoxicity assays demonstrated that only PTK7-CAR2 modified T cells retained effective through multiple rounds of tumor challenge. Using in vivo xenograft models of lung cancers with different expression levels of PTK7, systemic delivery of PTK7-CAR2 modified T cells significantly prevented tumor growth and prolonged overall survival of mice. Altogether, our results support PTK7 as a therapeutic target suitable for CAR T-cell therapy that could be applied for lung cancers and many other solid cancers with PTK7 overexpression.

Fructose-1,6-Bisphosphate Aldolase B Depletion Promotes Hepatocellular Carcinogenesis Through Activating Insulin Receptor Signaling and Lipogenesis.

BACKGROUND AND AIMS: Insulin receptor (IR) transduces cell surface signal through phosphoinositide 3-kinase (PI3K)-AKT pathways or translocates to the nucleus and binds to the promoters to regulate genes associated with insulin actions, including de novo lipogenesis (DNL). Chronic activation of IR signaling drives malignant transformation, but the underlying mechanisms remain poorly defined. Down-regulation of fructose-1,6-bisphosphate aldolase (ALDO) B in hepatocellular carcinoma (HCC) is correlated with poor prognosis. We aim to study whether and how ALDOB is involved in IR signaling in HCC. APPROACH AND RESULTS: Global or liver-specific ALDOB knockout (L-ALDOB-/- ) mice were used in N-diethylnitrosamine (DEN)-induced HCC models, whereas restoration of ALDOB expression was achieved in L-ALDOB-/- mice by adeno-associated virus (AAV). 13 C6 -glucose was employed in metabolic flux analysis to track the de novo fatty acid synthesis from glucose, and nontargeted lipidomics and targeted fatty acid analysis using mass spectrometry were performed. We found that ALDOB physically interacts with IR and attenuates IR signaling through down-regulating PI3K-AKT pathways and suppressing IR nuclear translocation. ALDOB depletion or disruption of IR/ALDOB interaction in ALDOB mutants promotes DNL and tumorigenesis, which is significantly attenuated with ALDOB restoration in L-ALDOB-/- mice. Notably, attenuated IR/ALDOB interaction in ALDOB-R46A mutant exhibits more significant tumorigenesis than releasing ALDOB/AKT interaction in ALDOB-R43A, whereas knockdown IR sufficiently diminishes tumor-promoting effects in both mutants. Furthermore, inhibiting phosphorylated AKT or fatty acid synthase significantly attenuates HCC in L-ALDOB-/- mice. Consistently, ALDOB down-regulation is correlated with up-regulation of IR signaling and DNL in human HCC tumor tissues. CONCLUSIONS: Our study reports a mechanism by which loss of ALDOB activates IR signaling primarily through releasing IR/ALDOB interaction to promote DNL and HCC, highlighting a potential therapeutic strategy in HCC.

Novel small molecule inhibitors of the transcription factor ETS-1 and their antitumor activity against hepatocellular carcinoma.

The transcription factor ETS-1 (E26 transformation specific sequence 1) is the key regulator for malignant tumor cell proliferation and invasion by mediating the transcription of the invasion/migration related factors, e.g. MMPs (matrix metalloproteinases). This work aims to identify the novel small molecule inhibitors of ETS-1 using a small molecule compound library and to study the inhibitors' antitumor activity against hepatocellular carcinoma (HCC). The luciferase reporter is used to examine the inhibition and activation of ETS-1's transcription factor activity in HCC cells, including a highly invasive HCC cell line, MHCC97-H, and five lines of patient-derived cells. The inhibition of the proliferation of HCC cells is examined using the MTT assay, while the invasion of HCC cells is examined using the transwell assay. The anti-tumor activity of the selected compound on HCC cells is also examined in a subcutaneous tumor model or intrahepatic tumor model in nude mice. The results show that for the first time, four compounds, EI1~EI-4, can inhibit the transcription factor activation of ETS-1 and the proliferation or invasion of HCC cells. Among the four compounds, EI-4 has the best activation. The results from this paper contribute to expanding our understanding of ETS-1 and provide alternative, the safer and more effective, HCC molecular therapy strategies.

Quantitative analysis of impurities in leucomycin bulk drugs and tablets: A high performance liquid chromatography-charged aerosol detection method and its conversion to ultraviolet detection method.

Toxic impurities were found in leucomycin and its preparation, however the content determination of impurities was challengeable due to the lacking of their reference standards. In this study, we developed high-performance liquid chromatography method coupled with charged aerosol detection (CAD) for the quantification of related substance of leucomycin (kitasamycin) bulk drugs and tablets, however, the CAD was not yet popular. In order to carry out quantitation work conveniently in the laboratory without CAD instruments, a high-performance liquid chromatography method coupled with ultraviolet (UV) detection was developed with the assistant of the HPLC-CAD results. The relative response of impurities on CAD chromatogram was used for guiding the establishment of HPLC-UV method, which could achieve the quantitation task in the absence of impurity reference standards. The developed HPLC-UV method was validated according to the ICH guideline and showed good precision, reproducibility and linearity with determination coefficient higher than 0.9999. The limit of detection and quantitation were 0.3 and 0.5 µg mL-1, respectively. The recoveries were 92.9 %-101.5 % at the spiked concentration levels of 0.1 %, 0.8 %, 1.0 and 1.2 % with relative standard deviations (RSDs, n = 3) lower than 2.0 %. Finally, the developed HPLC-CAD and -UV methods were compared by the determination of impurities in several batches of leucomycin bulk drugs and tablets. The results demonstrated that the developed HPLC-UV method was simple and reliable. This study developed methods to quantify the related substance in leucomycin and tablets, and discussed a strategy of the conversion of HPLC-CAD method to HPLC-UV method. The developed methods could be considered for implementation into pharmacopeial monographs in the future.