Echinacoside regulates PI3K/AKT/HIF-1α/VEGF cross signaling axis in proliferation and apoptosis of breast cancer.

CONTEXT: Echinacoside (ECH) is a natural anti-cancer compound and is of great value in cancer treatment. However, the mechanism underlying this effect on breast cancer (BC) was unclear. OBJECTIVE: To explore the mechanism of ECH treating BC by network pharmacology and experimental validation. MATERIALS & METHODS: Several databases were searched to screen potential targets of ECH and obtain information on targets related to BC. STRING was applied to construct a Protein-protein interaction (PPI) network. DAVID was applied for Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. Gene Expression Profiling Interactive Analysis (GEPIA) was searched for the relationship between the expression profile and overall survival of major targets in normal breast and BC tissues. Finally, the results of network pharmacology analysis were validated by experiments. RESULTS: Seventeen targets of ECH overlapped with targets in BC. Ten hub targets were determined through PPI. By GO and KEGG analysis 15 entries and 25 pathways were obtained, in which phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), hypoxia inducible factor-1 (HIF-1) and vascular endothelial growth factor (VEGF) played greater roles. Validation of key targets in the GEPIA database showed that PIK3R1 and PIK3CD remained consistent with the results of the study. Experiments in vitro showed ECH inhibited proliferation, induced apoptosis and reduced mRNA levels and protein expression of PI3K, AKT, hypoxia inducible factor-1α (HIF-1α) and vascular endothelial growth factor A (VEGFA) in MCF-7 cells. Furthermore, experiments in vivo revealed that ECH significantly reduced tumor growth, promoted apoptosis and decreased the related mRNA levels and protein expression, suggesting ECH works on BC by regulating PI3K/AKT/HIF-1α/VEGF signaling pathway. DISCUSSION & CONCLUSION: In summary, ECH played an important role in anti-BC by regulating PI3K/AKT/HIF-1α/VEGF signaling pathway. Furthermore, ECH had multi-target and multi-pathway effects, which may be a promising natural compound for treating BC.

Environmental exposure and ecological risk of perfluorinated substances (PFASs) in the Shaying River Basin, China.

There have been concerns raised about the environmental effects of perfluoroalkyl substances (PFASs) because of their toxicity, widespread distribution, and persistence. Understanding the occurrences and ecological risk posed by PFASs is essential, especially for the short-chain replacements perfluorobutanoic acid (PFBA) and perfluorobutane sulfonic acid (PFBS), which are now becoming predominant PFASs. The lack of aquatic life criteria (ALC), however, prevents an accurate assessment of the ecological risks of PFBA and PFBS. This study thus investigated the occurrence of 15 PFASs at 29 sampling sites in Shaying River Basin (in China) systematically, conducted the toxicity tests of PFBA and PFBS on eight resident aquatic organisms in China, and derived the predicted non-effect concentration (PNEC) values for PFBA and PFBS for two environmental media in China. The results showed that the total PFASs concentrations (ΣPFASs) ranged from 5.07 to 20.32 ng/L (average of 10.95 ng/L) in surface water, whereas in sediment, ΣPFASs ranged from 6.46 to 20.05 ng/g (dw) (average of 11.51 ng/g). The presence of PFBS was the most prominent PFASs in both water (0.372-8.194 ng/L) and sediment (4.54-15.72 ng/g), demonstrating that short-chain substitution effects can be observed in watersheds. The PNEC values for freshwater and sediment were 6.60 mg/L and 8.30 mg/kg (ww), respectively, for PFBA, and 14.04 mg/L, 37.08 mg/kg (ww), respectively, for PFBS. Ecological risk assessment of two long-chain PFASs, perfluorooctanoic acid (PFOA) and perfluorooctane sulfonate (PFOS), and two short-chain PFASs, PFBA and PFBS, using the hazard quotient method revealed that Shaying River and other major River Basins in China were at risk of PFOS contamination. This study contributes to a better understanding of the presence and risk of PFASs in the Shaying River and first proposes the ALCs for PFBA and PFBS in China, which could provide important reference information for water quality standards.

[Effect of diosgenin on mTOR/FASN/HIF-1α/VEGFA expression in rats with non-alcoholic fatty liver disease].

The present study aimed to investigate the effect of diosgenin on mammalian target of rapamycin(mTOR), fatty acid synthase(FASN), hypoxia inducible factor-1α(HIF-1α), and vascular endothelial growth factor A(VEGFA) expression in liver tissues of rats with non-alcoholic fatty liver disease(NAFLD) and explore the mechanism of diosgenin on lipogenesis and inflammation in NAFLD. Forty male SD rats were divided into a normal group(n=8) fed on the normal diet and an experimental group(n=32) fed on the high-fat diet(HFD) for the induction of the NAFLD model. After modeling, the rats in the experimental group were randomly divided into an HFD group, a low-dose diosgenin group(150 mg·kg~(-1)·d~(-1)), a high-dose diosgenin group(300 mg·kg~(-1)·d~(-1)), and a simvastatin group(4 mg·kg~(-1)·d~(-1)), with eight rats in each group. The drugs were continuously given by gavage for eight weeks. The levels of triglyceride(TG), total cholesterol(TC), low-density lipoprotein cholesterol(LDL-C), alanine transaminase(ALT), and aspartate transaminase(AST) in the serum were detected by the biochemical method. The content of TG and TC in the liver was detected by the enzyme method. Enzyme-linked immunosorbent assay(ELISA) was used to measure interleukin 1ß(IL-1ß) and tumor necrosis factor α(TNF-α) in the serum. Lipid accumulation in the liver was detected by oil red O staining. Pathological changes of liver tissues were detected by hematoxylin-eosin(HE) staining. The mRNA and protein expression levels of mTOR, FASN, HIF-1α, and VEGFA in the liver of rats were detected by real-time fluorescence-based quantitative polymerase chain reaction(PCR) and Western blot, respectively. Compared with the normal group, the HFD group showed elevated body weight and levels of TG, TC, LDL-C, ALT, AST, IL-1ß, and TNF-α(P<0.01), increased lipid accumulation in the liver(P<0.01), obvious liver steatosis, up-regulated mRNA expression levels of mTOR, FASN, HIF-1α, and VEGFA(P<0.01), and increased protein expression levels of p-mTOR, FASN, HIF-1α, and VEGFA(P<0.01). Compared with the HFD group, the groups with drug treatment showed lowered body weight and levels of TG, TC, LDL-C, ALT, AST, IL-1ß, and TNF-α(P<0.05, P<0.01), reduced lipid accumulation in the liver(P<0.01), improved liver steatosis, decreased mRNA expression levels of mTOR, FASN, HIF-1α, and VEGFA(P<0.05, P<0.01), and declining protein expression levels of p-mTOR, FASN, HIF-1α, and VEGFA(P<0.01). The therapeutic effect of the high-dose diosgenin group was superior to that of the low-dose diosgenin group and the simvastatin group. Diosgenin may reduce liver lipid synthesis and inflammation and potentiate by down-regulating the mTOR, FASN, HIF-1α, and VEGFA expression, playing an active role in preventing and treating NAFLD.

Shuangyu Tiaozhi decoction alleviates non-alcoholic fatty liver disease by improving lipid deposition, insulin resistance, and inflammation in vitro and in vivo.

Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases worldwide. Our previous studies have found that Shuangyu Tiaozhi Decoction (SYTZD) could produce an improvement in NAFLD-related indicators, but the underlying mechanism associated with this improvement remains unclear. The study aimed to investigate the potential mechanism of SYTZD against NAFLD through network pharmacology and experimental verification. The components of SYTZD and SYTZD drug containing serum were analyzed using ultra-performance liquid chromatography to quadrupole/time-of-flight mass spectrometry (UPLC-Q/TOF-MS). Active components and targets of SYTZD were screened by the traditional Chinese medical systems pharmacology (TCMSP) and encyclopedia of traditional Chinese medicine (ETCM) databases. NAFLD-related targets were collected from the GeneCards and DisGeNET databases. The component-disease targets were mapped to identify the common targets of SYTZD against NAFLD. Protein-protein interaction (PPI) network of the common targets was constructed for selecting the core targets. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the core targets was performed using the database for annotation, visualization, and integrated discovery (DAVID) database. Furthermore, animal and cell models were constructed for validating the predictions of network pharmacology. Lipid accumulation, liver histopathology, insulin resistance, and core gene expression were measured by oil red O staining, hematoxylin and eosin staining, insulin tolerance test, real-time quantitative polymerase chain reaction, and Western blotting, respectively. Two components and 22 targets of SYTZD against NAFLD were identified by UPLC-Q/TOF-MS and relevant databases. PPI analysis found that ESR1, FASN, mTOR, HIF-1α, VEGFA, and GSK-3ß might be the core targets of SYTZD against NAFLD, which were mainly enriched in the thyroid hormone pathway, insulin resistance pathway, HIF-1 pathway, mTOR pathway, and AMPK pathway. Experimental results revealed that SYTZD might exert multiple anti-NAFLD mechanisms, including improvements in lipid deposition, inflammation, and insulin resistance. SYTZD treatment led to decreases in the lipid profiles, hepatic enzyme levels, inflammatory cytokines, and homeostatic model assessment for insulin resistance (HOMA-IR). SYTZD treatment affected relative mRNA and protein levels associated with various pathways. Our findings reveal that SYTZD could alleviate NAFLD through a multi-component, multi-target, and multi-pathway mechanism of action.

Migraine Is Associated With High Risk of Erectile Dysfunction: A Systematic Review and Cumulative Analysis.

BACKGROUND: Migraine, a common chronic primary headache, has been found to be associated with a high risk of erectile dysfunction (ED). AIM: The present study aims to summarize all the evidence related to this topic and demonstrate a quantified result on the association between migraine and ED, which has not been reported in the literature. METHODS: MEDLINE, Excerpta Medica Database, and Cochrane Library were systematically searched for identifying the eligible studies (2000-2021). This study was registered in the PROSPERO (ID: CRD42021248013). OUTCOMES: The combined effects were synthesized with the relative risks (RR) or standard mean differences (SMD) with 95% confidence intervals (CI). RESULTS: 6 trials with a total of 51,657 participants were included, of which 6,175 were men with migraine. The pooled analysis indicated that migraine was associated with a significantly higher risk of ED as compared to the non-migraine general population (RR = 1.63, 95%CI: 1.34 to 2.0, P < .001). Consistently, men with migraine have a significantly lower IIEF-5 score than healthy controls (SMD = -3.64, 95%CI: -6.4 to -0.89, P = .01). Stratification analysis on the mean age indicated that the association between migraine and ED was much stronger in the migraine patients with age < 40 years (RR = 32.29, 95% CI: 6.41-162.64, P < .001; I2 = 0.0 %, P = .837) than in those with age > 40 years (RR = 1.75, 95% CI: 1.11-2.78, P = .017; I2 = 89.2%, P = .002). Sensitivity analysis indicated that no single study had dominated the combined RR and the heterogeneity. CLINICAL IMPLICATIONS: ED is a common disease among migraine men, especially those patients whose age is under 40 years old. It shows a 32-fold increased risk of ED compared to the healthy controls. Migraine-induced ED may be correlated with multiple factors, that is, chronic illnesses, chronic pain, and psychosocial causes (like anxiety and depression). Since phosphodiesterase-5 inhibitors (ie, sildenafil) might induce or exacerbate migraine, thus it is not recommended to prescribe these drugs for patients with migraine-mediated ED. CONCLUSION: The present study provides evidence that migraine is associated with a significantly high risk of ED, especially in those aged < 40 years. The pathophysiological mechanisms of this action deserve further study. He W, Yang Y, Liang H, et al. Migraine Is Associated With High Risk of Erectile Dysfunction: A Systematic Review and Cumulative Analysis. J Sex Med 2022;19:430-440.

Development and Validation of a Predictive Scoring System for In-hospital Death in Patients With Intra-Abdominal Infection: A Single-Center 10-Year Retrospective Study.

Objective: To develop and validate a scoring system to predict the risk of in-hospital death in patients with intra-abdominal infection (IAI). Materials and Methods: Patients with IAI (n = 417) treated at our hospital between June 2010 and May 2020 were retrospectively reviewed. Risk factors for in-hospital death were identified by logistic regression analysis. The regression coefficients of each risk factor were re-assigned using the mathematical transformation principle to establish a convenient predictive scoring system. The scoring system was internally validated by bootstrapping sample method. Results: Fifty-three (53/417, 12.7%) patients died during hospitalization. On logistic regression analysis, high APACHE II score (P = 0.012), pneumonia (P = 0.002), abdominal surgery (P = 0.001), hypoproteinemia (P = 0.025), and chronic renal insufficiency (P = 0.001) were independent risk factors for in-hospital death. On receiver operating characteristic curve analysis, the composite index combining these five risk factors showed a 62.3% sensitivity and 80.2% specificity for predicting in-hospital death (area under the curve: 0.778; 95% confidence interval: 0.711-0.845, P < 0.001). The predictive ability of the composite index was better than that of each independent risk factor. A scoring system (0-14 points) was established by re-assigning each risk factor based on the logistic regression coefficient: APACHE II score (10-15 score, 1 point; >15 score, 4 points); pneumonia (2 points), abdominal surgery (2 points), hypoproteinemia (2 points), and chronic renal insufficiency (4 points). Internal validation by 1,000 bootstrapping sample showed relatively high discriminative ability of the scoring system (C-index = 0.756, 95% confidence interval: 0.753-0.758). Conclusions: The predictive scoring system based on APACHE II score, pneumonia, abdominal surgery, hypoproteinemia, and chronic renal insufficiency can help predict the risk of in-hospital death in patients with IAI.