Potential anti-liver cancer targets and mechanisms of kaempferitrin based on network pharmacology, molecular docking and experimental verification.

AIM: Kaempferitrin is an active component in Chenopodium ambrosioides, showing medicinal functions against liver cancer. This study aimed to identify the potential targets and pathways of kaempferitrin against liver cancer using network pharmacology and molecular docking, and verify the essential hub targets and pathway in mice model of SMMC-7721 cells xenografted tumors and SMMC-7721 cells. METHODS: Kaempferitrin therapeutical targets were obtained by searching SwissTargetPrediction, PharmMapper, STITCH, DrugBank, and TTD databases. Liver cancer specific genes were obtained by searching GeneCards, DrugBank, TTD, OMIM, and DisGeNET databases. PPI network of "kaempferitrin-targets-liver cancer" was constructed to screen the hub targets. GO, KEGG pathway and MCODE clustering analyses were performed to identify possible enrichment of genes with specific biological subjects. Molecular docking and molecular dynamics simulation were employed to determine the docking pose, potential and stability of kaempferitrin with hub targets. The potential anti-liver cancer mechanisms of kaempferitrin, as predicted by network pharmacology analyses, were verified by in vitro and in vivo experiments. RESULTS: 228 kaempferitrin targets and 2186 liver cancer specific targets were identified, of which 50 targets were overlapped. 8 hub targets were identified through network topology analysis, and only SIRT1 and TP53 had a potent binding activity with kaempferitrin as indicated by molecular docking and molecular dynamics simulation. MCODE clustering analysis revealed the most significant functional module of PPI network including SIRT1 and TP53 was mainly related to cell apoptosis. GO and KEGG enrichment analyses suggested that kaempferitrin exerted therapeutic effects on liver cancer possibly by promoting apoptosis via p21/Bcl-2/Caspase 3 signaling pathway, which were confirmed by in vivo and in vitro experiments, such as HE staining of tumor tissues, CCK-8, qRT-PCR and Western blot. CONCLUSION: This study provided not only insight into how kaempferitrin could act against liver cancer by identifying hub targets and their associated signaling pathways, but also experimental evidence for the clinical use of kaempferitrin in liver cancer treatment.

Extracellular vesicle-packaged ILK from mesothelial cells promotes fibroblast activation in peritoneal fibrosis.

Progressive peritoneal fibrosis and the loss of peritoneal function often emerged in patients undergoing long-term peritoneal dialysis (PD), resulting in PD therapy failure. Varieties of cell-cell communications among peritoneal cells play a significant role in peritoneal fibrogenesis. Extracellular vesicles (EVs) have been confirmed to involve in intercellular communication by transmitting proteins, nucleic acids or lipids. However, their roles and functional mechanisms in peritoneal fibrosis remain to be determined. Using integrative analysis of EV proteomics and single-cell RNA sequencing, we characterized the EVs isolated from PD patient's effluent and revealed that mesothelial cells are the main source of EVs in PD effluent. We demonstrated that transforming growth factor-ß1 (TGF-ß1) can substitute for PD fluid to stimulate mesothelial cells releasing EVs, which in turn promoted fibroblast activation and peritoneal fibrogenesis. Blockade of EVs secretion by GW4869 or Rab27a knockdown markedly suppressed PD-induced fibroblast activation and peritoneal fibrosis. Mechanistically, injured mesothelial cells produced EVs containing high level of integrin-linked kinase (ILK), which was delivered to fibroblast and activated them via p38 MAPK signalling pathway. Clinically, the expression of ILK was up-regulated in fibrotic peritoneum of patients undergoing long-term PD. The percentage of ILK positive EVs in PD effluent correlated with peritoneal dysfunction and the degree of peritoneal damage. Our study highlights that peritoneal EVs mediate communications between mesothelial cells and fibroblasts to initiate peritoneal fibrogenesis. Targeting EVs or ILK could provide a novel therapeutic strategy to combat peritoneal fibrosis.

Kaempferitrin, a major compound from ethanol extract of Chenopodium ambrosioides, exerts antitumour and hepatoprotective effects in the mice model of human liver cancer xenografts.

OBJECTIVES：: This study aimed to investigate antitumour effect and possible toxicity of kaempferitrin, the major compound from ethanol extract of Chenopodium ambrosioides, in the mice model of human liver cancer xenografts. METHODS：: Forty mice bearing SMMC-7721 cells xenografts were divided into control group (not treated) and three groups orally administered with ethanol extract of C. ambrosioides, kaempferol (positive control) and kaempferitrin for 30 days. Antitumour effect was evaluated by measurement of tumour growth, histological examinations of tumours, flow cytometry detection of splenic CD19+ B lymphocytes and CD161+ Natural Killer cells, biochemical measurements of serum levels of tumour necrosis factor-α, interleukin-6, interferon-γ, malonaldehyde, 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azinobis-(3-ethylbenz thiazoline-6-sulphonate) radicals. Toxicity was evaluated by histological examinations of livers and measurements of serum levels of aspartate transaminase, alanine transaminase, total bilirubin, direct bilirubin, malonaldehyde and hepatic malonaldehyde level. KEY FINDINGS: Kaempferitrin significantly (P < 0.05) decreased tumour volume, mass and cell number. Antitumour effect was due to induction of tumour cells necrosis and apoptosis, stimulation of splenic B lymphocytes, decreases of radicals and malonaldehyde. Kaempferitrin did not change liver structure, and decreased serum levels of transaminases, bilirubin, malonaldehyde and hepatic malonaldehyde level. CONCLUSIONS: Kaempferitrin exerts antitumour and hepatoprotective effects.

Proteomic Characterization of Peritoneal Extracellular Vesicles in a Mouse Model of Peritoneal Fibrosis.

Peritoneal fibrosis progression is regarded as a significant cause of the loss of peritoneal function, markedly limiting the application of peritoneal dialysis (PD). However, the pathogenesis of peritoneal fibrosis remains to be elucidated. Tissue-derived extracellular vesicles (EVs) change their molecular cargos to adapt the environment alteration, mediating intercellular communications and play a significant role in organ fibrosis. Hence, we performed, for the first time, four-dimensional label-free quantitative liquid chromatography-tandem mass spectrometry proteomic analyses on EVs from normal peritoneal tissues and PD-induced fibrotic peritoneum in mice. We demonstrated the alterations of EV concentration and protein composition between normal control and PD groups. A total of 2339 proteins containing 967 differentially expressed proteins were identified. Notably, upregulated proteins in PD EVs were enriched in processes including response to wounding and leukocyte migration, which participated in the development of fibrosis. In addition, EV proteins of the PD group exhibited unique metabolic signature compared with those of the control group. The glycolysis-related proteins increased in PD EVs, while oxidative phosphorylation and fatty acid metabolism-related proteins decreased. We also evaluated the effect of cell-type specificity on EV proteins, suggesting that mesothelial cells mainly cause the alterations in the molecular composition of EVs. Our study provided a useful resource for further validation of the key regulator or therapeutic target of peritoneal fibrosis.

Integrated metabolomic and transcriptomic analyses revealed metabolite variations and regulatory networks in Cinnamomum cassia Presl from four growth years.

To understand the mechanism of the dynamic accumulation of active ingredients in Cinnamomum cassia Presl, metabolomic and transcriptomic analyses of 5~8 years old C. cassia were performed. A total of 72 phenylpropanoids, 146 flavonoids, and 130 terpenoids showed marked changes. Most phenylpropanoids and flavonoids showed markedly higher abundances in 6-year-old C. cassia than in others, which was related to the higher expression of genes that synthesize and regulate phenylpropanoids and flavonoid. We identified transcription factors (TFs) and genes involved in phenylpropanoids and flavonoids synthesis and regulation through co-expression network analyses. Furthermore, most of the terpenoids in 5-year-old C. cassia showed markedly higher abundances than in others, which was due to the differentially expressed genes upstream of the terpenoids pathway. The results of our study provide new insights into the synthesis and accumulation of phenylpropanoid, flavonoids and terpenoids in C. cassia at four growth stages.

Development, Reliability, and Validity of the Home Blood Pressure Monitoring Adherence Scale for Patients with Chronic Kidney Disease.

BACKGROUND: Home blood pressure monitoring helps patients with chronic kidney disease to improve blood pressure control and can predict cardiovascular events, renal function progress, and risk of death. Few instruments are available to assess patient adherence to home blood pressure monitoring. OBJECTIVE: The aim of the study was to develop an instrument to evaluate home blood pressure monitoring adherence in patients with chronic kidney disease and test its reliability and validity. METHODS: An item pool was formed for the Home Blood Pressure Monitoring Adherence Scale by literature review. Patients with chronic kidney disease (n = 436) were surveyed to assess item selection and examine item reliability and validity. Scale reliability was evaluated using internal, split-half, and test-retest reliability, while validity was assessed according to content, construct, and criterion validity. RESULTS: The scale comprising eight items was formed from the item pool and item selection. Cronbach's α was 0.906, split-half reliability was 0.947, and test-retest reliability was 0.716. Item-level and scale-level (both universal agreement and average) content validity indices were 1.00. According to the Self-Efficacy for Managing Chronic Disease 6-item Scale, criterion validity for our scale was 0.251. Exploratory factor analysis extracted one factor and the cumulative variance contribution rate was 61.568%. Confirmatory factor analysis showed the model fit well (Χ 2=50.125, df=17, Χ 2 /df=2.949, root mean square error of approximation=0.095, confirmatory fit index=0.970). CONCLUSION: The scale has good reliability and validity for patients with chronic kidney disease, representing an efficient instrument for clinical assessment of home blood pressure monitoring adherence.

Prognostic value of pulmonary hypertension in pre-dialysis chronic kidney disease patients.

BACKGROUND: Pulmonary hypertension is common in chronic kidney disease (CKD) patients. However, the prognostic value of pulmonary hypertension in Chinese predialytic CKD patients is rarely reported. We evaluated the relevant factors and prognostic value of pulmonary hypertension in CKD patients. METHODS: This retrospective cohort study enrolled 1092 predialytic patients from The Third Affiliated Hospital of Sun Yat-Sen University from May 1st, 2011, to December 31st, 2016. Data of interest were retrieved from electronic medical records. Pulmonary hypertension was defined as pulmonary arterial systolic pressure (PASP) ≥ 35 mmHg by echocardiology. All participants were followed from the date of the first echocardiography examination. The primary endpoints were all-cause mortality and cardiovascular mortality. The secondary endpoint was end-stage renal disease (ESRD) defined as starting renal replacement therapy. RESULTS: The prevalence of pulmonary hypertension was 15.9% in the study population. For CKD stage 1, 2, 3a, 3b, 4 and 5, the prevalence was 6.0%, 9.6%, 17.2%, 13.3%, 20.7% and 26.6%, respectively. Older age, lower left ventricular ejection fraction, anemia and higher pulse pressure were independently associated with pulmonary hypertension in CKD patients. In multivariate Cox regression analysis, pulmonary hypertension was the independent risk factor for cardiovascular mortality, but not of all-cause mortality and ESRD. CONCLUSIONS: Pulmonary hypertension is not rare in early CKD patients. Patients with older age, anemia, higher pulse pressure and compromised heart function were more likely to comorbid pulmonary hypertension. Pulmonary hypertension maybe a sign of worse cardiovascular outcome in CKD patients.

Magnesium sulfate for acute traumatic brain injury.

BACKGROUND: The purpose of this systematic review was to evaluate the effect of magnesium sulfate in the treatment of acute traumatic brain injury. MATERIALS AND METHODS: A systematic search of ClinicalTrials.gov, the Cochrane Library database, EMBASE, MEDLINE, Web of Science, and the World Health Organization trial registry, plus manual searches of gray literature, was undertaken in April 2013. Two reviewers independently extracted the data with a predefined data extraction form. RevMan 5 software was used to synthesize data and calculate the risk ratio for mortality with the 95% confidence interval. For the Glasgow Outcome Scale and posttreatment Glasgow Coma Scale data, the weighted mean difference was calculated with the 95% confidence interval. RESULTS: A total of 8 randomized controlled trials with a total of 786 patients were included. Meta-analysis showed that there was no significant difference between the groups for mortality. The Glasgow Outcome Scale of the treatment group was higher than that of the control group, although the significance was borderline. The Glasgow Coma Scale score change posttreatment was significantly higher than that of the control. CONCLUSIONS: The present meta-analysis of existing randomized controlled trials does not identify a significant beneficial effect in the mortality of traumatic brain injury patients; however, it suggests that magnesium sulfate shows a tendency to improve the Glasgow Outcome Scale and Glasgow Coma Scale scores, which is a promising result for traumatic brain injury therapy. Further effort is necessary to explore which subgroup of traumatic brain injury patients could benefit from magnesium sulfate.

Effect of methyl jasmonate on the anthocyanin content and antioxidant activity of blueberries during cold storage.

BACKGROUND: The effects of postharvest methyl jasmonate (MeJA) fumigation on total soluble solids (TSS), titratable acidity (TA), ascorbic acid, total phenolic content (TPC), total monomeric anthocyanins (TMAs), individual anthocyanins and antioxidant activity of blueberries stored at 1 °C for 28 days were evaluated. Prior to storage, the blueberries were fumigated with 0.05 mmol L(-1) MeJA for 12 h. Control blueberries were subjected to the same conditions but were not exposed to MeJA. RESULTS: MeJA treatment had no adverse effect on TSS and TA and inhibited the decrease in ascorbic acid during storage. MeJA treatment induced an enhancement in TPC on day 21; TPC decreased thereafter. Similarly, a significant increase in TMAs and individual anthocyanins was observed 21 days after MeJA treatment. TPC, TMAs and individual anthocyanins increased in control fruits on day 7 and decreased thereafter. Moreover, MeJA treatment maintained higher levels of antioxidant activity during the entire storage period. CONCLUSION: These results suggest that cold storage enhances TPC, TMAs and individual anthocyanin content during short-term storage. However, postharvest application of MeJA to blueberries enhances TPC, TMAs and individual anthocyanin content during long-term storage.

Free fatty acid secretion by an engineered strain of Escherichia coli.

Although most bacteria produce fatty acids (FA), few secrete free FAs into the culture media. Over-expression of two FA thioesterases, TesA and AtFatA, facilitated both total and FFA production in a recombinant strain of Escherichia coli. When these thioesterases were expressed in a fadD and fadL double-deletion strain, a further enhancement of FFA secretion was observed. These results support a simple diffusion mechanism for FA transport. In addition, the ATP-binding cassette transporter protein, MsbA, also increased the concentration of FFAs in the culture. The final strain produced 110 mg FFA/l, about 33 % of the total FAs being produced. Our findings support a diffusion mechanism for FA transport.